Raffles Institution Nov 2007 (H2 Biology) Paper 2 (for 9744 syllabus)

Nov 2007 H2 Bio Paper 2

N07P2Q1
1a) Fig. 1.1 shows the main structural features of a mature (infective) human
immunodeficiency virus (HIV).
Identify the molecules labelled A, B and C. [3]
A (Viral) RNA; R: Genome (because it is not a molecule)
B Reverse transcriptase (between integrase and reverse transcriptase, choose
the latter)
C Capsid protein/coat protein / capsomere (R:capsid)

Comments:
Given a choice between reverse transcriptase and integrase, choose reverse transcriptase.
What is wrong with this diagram – only one RNA molecule, (there should be two ss RNA
molecules); there should be another enzyme ie. integrase.

(b) Fig. 1.2 is an electron micrograph showing HIV being released from an infected cell.

Describe how HIV acquires the outer envelope. (4 lines) [3]

1. Viral glycoproteins e.g. gp 41 and gp120 are translated at rough endoplasmic reticulum*
from the mRNA of the provirus transcribed in the host.
2. These glycoproteins are then transported and inserted into the host cell membrane via
vesicles.
3. The nucleocapsid will then be assembled near the cell surface,
4. and it will bud off* from host cell membrane that is studded with viral glycoproteins to form a
new virus.
R: exocytosis 3 max

Comments:
Students can’t seem to focus on how ‘HIV acquires the outer envelope’ giving irrelevant
information.
Students should realize that the envelope is a modified host cell plasma membrane and so
synthesis of glycoprotein should be included.
Exocytosis is not the same as evagination/budding. What is the difference?

(c) After viruses have been released from an infected cell, HIV protease completes the
maturation of the viruses by cutting viral polyproteins to form the structural and
enzymatic proteins of the infective virus.
(i) Explain how HIV protease inhibitors work in the treatment of HIV infection. (4 lines)[3]
1. HIV protease inhibitors bind to the active site* of HIV protease and blocks the substrate
from binding;
2. so that the viral polyprotein cannot be cut into the necessary structural proteins and
enzymes;
3. (newly formed) virus not infective/ mature/ functional and unable to infect further cells;

Comments:
The question asks “how the inhibitor works” so an explanation of competitive inhibition is
required. Don’t simply rehash the question by saying “HIV protease inhibitors interferes
with the enzyme by inhibiting its activity.” How does it ‘interfere’ and how does it ‘inhibit’?
If possible, students should use relevant knowledge to infer and note that the ‘tunnel’ is
the active site and not use back the same term ‘tunnel’.
1
Raffles Institution Nov 2007 (H2 Biology) Paper 2 (for 9744 syllabus)

(ii) Describe what structural properties the inhibitor requires in order to perform its
function. (3 lines) [2]
1. similar shape and charge to viral polyprotein or complementary in shape and charge to
active site* of enzyme
2. does not break down easily / stable so can remain in active site*
Accept if used 3D conformation in place of ‘shape’
Accept also if candidate implied that molecule is ‘stable’

Comments:
Many couldn’t come up with a second point.
Although a permanent association between the inhibitor and enzyme is desirable, it
is not necessary.

N07P2Q2
(a) Fig. 1.1 shows a diagram of the fluid mosaic model of the cell surface membrane.

a) Explain why it is called a fluid mosaic. [3]

1. It is referred to as ‘fluid” because the cell membrane comprises of phospholipids*
and proteins* which are free to move laterally within a layer;
2. the phospholipids can flip flop from one layer to the other but this is a rare
occurrence.
3. It is referred to as ‘mosaic” because the random arrangement of the proteins
embedded amongst the phospholipid molecules resemble a mosaic pattern;

(b) The passage of most molecules through membranes is regulated by proteins. Fig. 1.2 is
a diagram showing the four main steps in the release of insulin from beta cells, which
involves three types of transmembrane proteins.

(i) Explain why transmembrane proteins are necessary for glucose, potassium ions and
calcium ions to pass through cell surface membrane. [3]
1. glucose is polar* while the ions are charged* and both are hydrophilic*;
2. Cell membranes have a hydrophobic core* making them impermeable to these solutes;
3. transmembrane transport proteins can be a channel or carrier that provides a
hydrophilic* channel through the membrane for the passage of the solutes;
4. because a transport protein is specific* to its own solute, different transport proteins are
needed for different solutes.

(ii) Describe how a change in the ATP-sensitive potassium channels will cause the calcium
channels to open. [3]
1. When ATP* levels are high, the ATP sensitive K+ channels close;
2. This leads to the build up of K+ ions inside the cell and the;
3. Subsequent depolarization* of the membrane opens the voltage gated Ca2+ channels.

(ii) Suggest why there are no channels for insulin release across the membrane. [2]
1. Insulin is a peptide hormone/protein that is too large to cross the membrane via a
channel (must have);
2. A channel with a big enough hydrophilic channel for insulin will allow many other
molecules to pass through as well.
3. so insulin is packaged in membrane-bound Golgi vesicles and released through process
of exocytosis* instead.

2
Raffles Institution Nov 2007 (H2 Biology) Paper 2 (for 9744 syllabus)

(c) Fig. 1.3 is an electronmicrograph showing release of insulin from a beta cell in the pancreas.

(i) Describe how this release across the membrane occurs. [3]
1. Golgi / secretory vesicles* containing insulin will be transported towards the cell
membrane along the microtubules;
2. Golgi/secretory vesicles* will fuse* with the cell membrane;
3. Releasing insulin to the exterior of the cell by exocytosis*.

N07P2Q3

(a) DNA is replicated semi-conservatively.

State what is meant by semi-conservative replication. [3] (4 lines)

1. DNA unzips* and strands separate as hydrogen bonds* between bases break;
2. Both strands serve as templates* for the synthesis of new strand by complementary
base pairing;
3. New molecule of DNA has one new strand and one original strand;

Comments:
2 strands make up one molecule of DNA/DNA double helix. Be careful with the use of
strands and molecules. Wrong usage(error underlined): The daughter strand is made up of
a newly synthesized strand and parental strand.
Many failed to mention hydrogen bonds breaking.

(b) Fig. 3.1 is a diagram showing part of a bacterial chromosome with a single origin of DNA
replication and two replication forks which proceed in opposite directions until replication
of the entire chromosome is complete. This takes about twenty minutes.

Explain why replication only occurs in a 5’ to 3’direction. [2] (3 lines)

1. The active site* of DNA polymerase is complementary to and binds the free 3’ OH
group of the growing DNA strand;
2. DNA polymerase adds DNA nucleotides to this 3’ end as the free 3’ OH group is needed
for DNA polymerase to start polymerisation;
Comments:
Some candidates made no reference to active site.
Should specify that polymerase binds to the free 3’ OH group on the “growing DNA
strand”. A free 3’ OH is also found in free DNA nucleotides too but we are not referring to
this!
A number did not mention the second point. Binding of enzyme active site to the 3’ OH
alone does not explain the direction of extension of the strand. Students need to explain the
5’ 3’ extension of the DNA strand.

(c) State two ways in which the structural arrangement of DNA differs between a bacterial
chromosome and human chromosomes. [2] (2 lines each)

1. Human chromosome is linear* while bacteria chromosome is circular;

2. Human DNA is arranged in nucleosomes wrapped around histones while bacteria DNA
is naked;
3. Human DNA has introns/telomeres/centromeres not present in bacteria but bacteria
DNA has operons;
4. Human chromosomes occur in pairs while bacteria chromosomes occur singly;
3
Raffles Institution Nov 2007 (H2 Biology) Paper 2 (for 9744 syllabus)

Comments:
The key word is structural arrangement of DNA. Not the size. Not the shape. (BTW the
human chromosome is not normally X-shaped. It only appears that way during prophase of
cell division. Normally it appears linear like a single thread).
Don’t confuse plasmids with bacteria chromosomes. They are extra-chromosomal in
addition to chromosomes. Bacteria has a circular chromosome and in some cases may
possess plasmids.
When making comparative statements, give a positive counterpoint where possible.
e.g. Human chromosome is linear while bacteria chromosome is not.
Human chromosome is linear while bacteria chromosome is circular.
Bacteria DNA is also a double helix. Where did you get the idea that it can be single
stranded?! Only viruses can have single or double stranded DNA or RNA.

(d) Replication of the largest human chromosome with only a single origin of DNA
replication would theoretically take approximately 800 hours.
a. Suggest two reasons why replication of a human chromosome would take so long. [2] (2
lines each)
1. DNA in human chromosomes are very long containing many non-coding DNA;
2. DNA in humans are packaged into nucleosomes and can be very condensed requiring
time to uncoil;

Comments:
Don’t quote an isolated figure like 150million base pairs or for that matter 150 billion base
pairs. Is that your way of saying that the chromosome is very long? What figure is
considered very long?
Lagging strand is not laid down slower than the leading strand. The replication fork
consisting of both strands progress at the same pace! Lagging strand therefore does not
hold back replication.
For the few who mentioned about histone proteins, you need to elaborate how it actually
slows replication.

In actively dividing human cells the process takes only twelve hours.
b. Describe how DNA replication is speeded up in human chromosomes. [1] (3 lines)

1. Replication of DNA in humans involve multiple origins/sites of replication where many

DNA polymerases can replicate DNA simultaneously;

Comments:
“Describe” requires more that just stating the presence of multiple origins of replication. Few
mentioned that this allowed many DNA polymerases to work simultaneously.
[Total:10]
N07/P2/Q4
Li-Fraumeni syndrome is a rare inherited cancer syndrome caused by mutations in a tumour
supressor gene on chromosome 17, known as p53. Mutations in p53 confer an increased risk
for early onset breast cancer, childhood sarcoma, osteosarcoma, brain tumours, leukaemia and
adrenocortical carcinoma.

(a) State the meaning of the term gene mutation. [3]

1. Alteration / changes in the DNA nucleotide sequence(s).
2. Can be a deletion/insertion mutation* where one or several nucleotides are removed
from / added into a DNA nucleotide sequence;
3. Can be a substitution mutation* where one nucleotide is replaced by another;
4
Raffles Institution Nov 2007 (H2 Biology) Paper 2 (for 9744 syllabus)

4. Can be an inversion mutation*, where a segment of nucleotide sequences separates

from the allele and rejoins at the original position, but is inverted;

(b) Explain how a mutation in the p53 gene could affect the structure of its product. [3]
1. Substitution mutation*:
a) one nucleotide is replaced with a different one.
b) change in codon* in mRNA, and an amino acid with a R group* of different
chemical property is coded for;
c) results in changes in interactions involved in maintaining tertiary structure e.g. ionic,
covalent/disulfide, hydrogen, hydrophobic interactions, lead to alteration of 3D
conformation of protein;
OR

2. Frameshift mutation*:
a) Due to deletion / insertion of nucleotides not in multiples of 3;
b) result in ribosomes reading mRNA template as incorrect triplets during translation /
alteration of reading frame;
c) all codons downstream of point of mutation will be read incorrectly, produce a
different sequence and number of amino acids;
d) results in non-functional protein / pre-mature termination of polypeptide should a stop
codon be generated;
OR
3. Addition or deletion mutation*:
a) Due to insertion / deletion of nucleotides not in multiples of 3;
b) result in frameshift of the codons on mRNA / alteration of reading frame;
c) all codons downstream of point of mutation will be read incorrectly, produce a
polypeptide with a different sequence and number of amino acids;
d) results in non-functional protein / pre-mature termination of polypeptide should a stop
codon be generated;

(c) Suggest why loss of function mutation in a tumour suppressor gene results in a recessive
allele. [2]
1. Loss of function mutation gives rise to a non-functional /absence of p53 protein;
2. Effect can be masked by a presence of normal dominant allele (functional copy of gene)
that will result in sufficient copies of the normal protein being synthesized to exert effect;
Or
Cancer will only manifest when both copies of tumour suppressor gene are mutated;

Comments:
Many students can explain why it is recessive but are not able to link it to the non-functional
protein produced. When we discuss about recessive and dominant, make reference to the
phenotype and only the gene products can affect the phenotype directly. The gene simply
carries the information that encodes the gene product. They don’t directly affect phenotype.
e.g. ‘defective gene is unable to restrict cell growth and proliferation” sounds strange.

(d) Fig 4.1 shows the protein produced by the normal p53 gene (p53 protein) attached to DNA.

Suggest why the p53 protein only binds to certain parts of the DNA molecule. [3]
1. DNA-binding domain of p53 protein;
2. has a conformation and charge* complementary* to specific parts of DNA molecule;
3. which has a specific sequence of bases and hence a specific conformation; (major and
minor grooves);
5
Raffles Institution Nov 2007 (H2 Biology) Paper 2 (for 9744 syllabus)

Comments:
Question is asking about specificity of DNA binding and not the type of genes they activate.
Why then do they show the figure of a protein wrapped around the DNA?
p53 does not go around scanning random segments of DNA for damage. It does not bind
damaged DNA. Other proteins do it and then once DNA damage is detected, these proteins
activate p53 which in turn activate genes responsible for DNA repair.
Few mentioned presence of a DNA binding domain and how its shape is complementary to
control elements.
Many simply said that “since it is a specific transcription factor, therefore it binds to specific
parts of DNA and not just any other part.” This statement doesn’t explain its specificity
which is what is required in this question.
p53 is NOT AN ENZYME so has no active site. It is a transcription factor. DNA-binding site
is not the same as active site. There are many different sites for different purposes. Be able
to distinguish them!
H3 Qn:What would you expect the common mutations on p53 to be? – Basic amino acid

(e) Fig 4.2 shows the structure of the core domain of the p53 protein with the six most frequently
mutated amino acids marked.

Suggest why some mutations of the p53 gene have no effect on the activity of the p53
protein. [4]
1. Genetic code is degenerate* where more than one type of codon* code for the same
amino acid;
mutations at 3rd nucleotide of codon still result in a codon coding for same amino acid;
OR
2. Mutation results in a codon coding for an amino acid, with an R group having similar
chemical properties to original amino acid;
OR
3. Mutation in the introns, which is spliced out and not translated, hence no change in
amino acid sequence;
OR
4. Mutation affects regions that are away from DNA binding site;

5. No change in conformation of DNA-binding site of p53;

6. Hence p53 still to able to bind to same specific regions on the DNA;

N07P2Q5
Fig. 5.1 shows comb shape in chickens.

(a) The genotype R-pp gives a rose comb, rrP- gives a pea comb, R-P- gives a walnut comb
and rrpp gives a single comb. The dash(-) indicates the presence of either the dominant or
recessive allele. List all the genotypes which will produce a walnut phenotype.[1]

RRPP, RRPp, RrPP and RrPp.

(b) Bateson and Punnett in a classic experiment, showing gene interactions, crossed chickens
with rose combs (Wyandotte pure line) with chickens with pea combs (Brahama pure line).
All the resulting F1 offspring differed from both the parents and had walnut combs.
When pairs of these F1 were allowed to interbreed they produced the resulting F2 ratio in
offspring:
9 walnut comb : 3 rose comb : 1 single comb : 3 pea comb
6
Raffles Institution Nov 2007 (H2 Biology) Paper 2 (for 9744 syllabus)

Draw a genetic diagram to explain both these crosses. [5]

Parental phenotype rose comb X pea comb
Parental genotype RRp’p’ rrPP
Rp’ rP
Gametes
F1 genotype RrPp’
F1 phenotype All Walnut (new phenotype)
Self-cross of F1 generation RrPp’ X RrPp’
F2

Gametes
RP Rp’ rP rp’

RRPP RRPp’ RrPP RrPp’

RP
walnut walnut walnut walnut
RRPp’ RRp’p’ RrPp’ Rrp’p’
Rp’
walnut rose walnut rose
RrPP RrPp’ rrPP rrPp’
rP
walnut walnut pea pea
RrPp’ Rrp’p’ rrPp’ rrp’p’
rp’
walnut rose pea single

F2 genotypes 9 R_P_ : 3 R_p’p’ : 3 rrP_ : 1 rrp’p’

F2 phenotypic ratio 9 walnut : 3 rose : 3 pea : 1 single

(c) A walnut comb chicken is crossed with a rose comb one. All the progeny are walnut combed.
State the two possible pairs of parental genotypes that would produce these progeny.
1. _________ x _________ 2. _________ x _________ [2]

RrPP x RRpp, RRPP x Rrpp or RRPP x RRpp

N07P2Q6
Fig. 6.1 is an electron micrograph of part of a plant cell.

(a) Name the structures P to R. [3]

P : nucleus
Q : nuclear envelope, (Nuclear membrane: not acceptable)
R : cell (surface)/ plasma membrane

(b) Suggest why plant cells with chloroplasts also contain mitochondria. [3]

1. Mitochondria are the sites for aerobic cellular respiration producing ATP;
2. ATP produced by mitochondria released for use in cytosol;
3. ATP from mitochondria is used for active transport/ macromolecule synthesis
(including DNA replication, transcription and translation) / formation of cytoskeleton
in cell division/vesicle movement etc;
4. However, ATP produced in chloroplasts during the light dependent stage is
subsequently used in the light independent stage of photosynthesis and is not
available for use for active processes in the cytosol;

7
Raffles Institution Nov 2007 (H2 Biology) Paper 2 (for 9744 syllabus)

(c) State 2 ways in which mitochondria and chloroplasts are

(i) similar in structure; [2]

1. Both are bound by a double membrane;

2. Inner membrane encloses a fluid-filled cavity which contains 70S ribosomes, circular
DNA strands and enzymes;
3. Electron transport chain found in the internal membranes of both organelles;
4. Stalked particles/ATP synthase are found in both organelles;

(ii) different in structure. [2]

Write in full sentences in answer. Do not use table in structured question.

Point of Chloroplast Mitochondria
Comparison
1. Size Larger (10µm in length and 5 Smaller (0.5µm wide and 2.5
µm in diameter) µm long)
2. Shape Lens shaped Generally spherical or rod
shaped
3. Inner Not folded, not arranged into Extensively folded as inner
membrane folds and do not contain portion of membrane contains
stalked particles stalked particles. Arranged
into folds known as cristae.
4.Granules/Grains Starch grains are present Numerous phosphate granules
present are present, no starch grains.
5.Internal Present in the form of stacks An internal membrane system
membrane of thylakoids (grana)* and is absent.
system intergranal lamella.
6.Location of Found on thylakoid Found on inner membrane.
stalked particles membranes.
7.Coloured Presence of photosynthetic There is an absence of
pigments pigments on thylakoid coloured pigments
membranes eg chlorophyll

(d) State how oxygen is involved in oxidative phosphorylation and in

photophosphorylation. [2]
1. In oxidative phosphorylation oxygen is the final electron acceptor as it combines with the
electrons and H+ to form water;
2. In photophosphorylation, oxygen is formed as a byproduct from photolysis;

N07P2Q7
The greater racket-tailed drongo, Dicrurus paradiseus, is an insect-eating bird found in tropical
broadleaved forests in Southern Asia from Kashmir, India and Sri Lanka east to Indonesia.
a) Complete Table 7.1 to show the classification of the greater racket-tailed drongo. [4]

kingdom > Animalia

> phylum Chordata
> class Aves
> order Passeriformes
family Dicruridae
genus > Dicrurus
> species Dicrurus paradiseus
8
Raffles Institution Nov 2007 (H2 Biology) Paper 2 (for 9744 syllabus)

Fig. 7.2 shows the geographic variation in the form of the crest among populations of the greater racket-
filled drongo.

b) Explain how the distinct phenotypic differences between the populations may have arisen. [5] 7L

1. There was geographical isolation* that prevented interbreeding and thus disrupted gene flow*
because there were many isolated islands and the broadleaved woodlands were not continuous;
2. Different regions had different environments, thus each sub-population was exposed to different
selection pressures*;
3. With variation in phenotypes among birds, natural selection will select for those best adapted to their
environment, who are more likely to survive, reproduce and passed on their alleles to the next
generation;
4. Allele frequencies change because of natural selection and genetic drift (which are random events
that are due to chance);
5. As the different populations evolve independently from each other, they accumulate different genetic
mutations and allele frequency changes over time and hence their distinct phenotypic differences;

R: reproductive isolation because they are still a species

c) Suggest why these populations of greater racket-tailed drongos are classified as a single species. [2]
3L
1. a group of organisms of the same species are capable of interbreeding* and producing fertile,
viable offspring*;
2. are reproductively isolated from other species;
3. have a common gene pool and same chromosome number;
4. usually have similar morphological, physiological and behavioural features;
[Total:11]

N07P2Q8
(a) Describe the main structural features of cellulose and collagen. [8]

1. Cellulose is a polysaccharide* made out of β glucose* residues linked together by β(1 4)

glycosidic bonds*
2. with alternate residues being inverted 180° with respect to its neighbours forming straight chains*;
3. Straight chains allow hydrogen bonding* to occur between hydroxyl groups of adjacent parallel
chains forming microfibrils*
4. Microfibrils are then bundled up to form macrofibrils*.

5. Collagen is a fibrous protein* made up of a chain of amino acids* joined by peptide bonds*.
6. The sequence is usually a repeating unit: glycine-X-Y* where X is usually proline and Y is usually
hydroxyproline.
7. Three loose helical chains coil and are held together by hydrogen bonds, forming the triple helix of
tropocollagen*.
8. Covalent bonds exist between staggered* tropocollagen molecules to form a collagen fibril
9. and bundles of collagen fibril form a collagen fibre.

(b) Explain how glucagon differs from glycogen. [8]

Point of glucagon glycogen
Comparison

a) Type of Glucagon is a globular protein. Glycogen is an extensively

macromolecule branched polysaccharide
(R: spherical/enzyme) made of helical chains.

9
Raffles Institution Nov 2007 (H2 Biology) Paper 2 (for 9744 syllabus)

b) Type of Monomers are amino acid Monomers consists of alpha

monomer residues* with different R glucose* only.
groups*.
c) Type of bonds Amino acid residues joined by Alpha glucose joined by alpha
between peptide bonds*. 1-4 glycosidic linkages* with
monomer branched alpha 1-6
glycosidic linkages*.

d) Number of Fixed number of amino acid per Variable number of glucose

monomer per molecule. per molecule.
molecule
e) Solubility in Soluble in water as it is globular Insoluble in water as it has a
water in structure. large molecular weight.

f) Synthesis Glucagon is produced by alpha Glycogen is synthesized in

cells in the pancreas in the islets liver and muscle cells when
of Langerhans. blood glucose is high.
g) Function Glucagon is a hormone/signal Glycogen provides an energy
protein that regulates blood store in muscle and liver.
glucose.
Each pt for pt comparison : 2 marks
Comments:
FULL sentences required. For internal exams, only -1, however, Cambridge will give ‘0’ if no
full sentences are written.
Some answered and compared differences only with respect to glucose
metabolism/regulation and lost many marks.
Good answers included points such as glycogen being a polysaccharide formed from a
chain of alpha glucose joined by alpha 1-4 with branched points with 1-6 glycosidic
linkages, while glucagon is a soluble globular protein made up of a chain of amino acids
joined by peptide bonds. Glycogen is insoluble in water whereas glucagon is soluble in
water and found in the blood plasma. Glycogen provides an energy store in muscle and
liver, whereas glucagon is a signal protein that regulates blood glucose, and is produced
by alpha cells in the pancreas in islets of Langerhans.

(c) Suggest why plant cells mainly store carbohydrates and animal cells mainly store lipids. [4]

1. Lipids contain more energy per unit mass compared to carbohydrates. Since animal are
more active than plants, more energy is required. Eg migrating birds will are less load
and more energy storing lipids compared to carbohydrates;
2. Lipids stored in fat cells of animals allow buoyancy, thermal insulation and protection of
vital organs;
3. Lipids provide more metabolic water compared to carbohydrates and hence useful for
storage in desert animals eg. camel;
4. Lipids in the form energy reserves are required in animals to hibernate through winter;
[Total : 20]

10
Raffles Institution Nov 2007 (H2 Biology) Paper 2 (for 9744 syllabus)

N07P2Q9

(a) Describe the main structural features of the lac operon. [8]

Diagram of lac operon (refer to diagram in subsequent points);

1. The lac operon is made of DNA and within it are 3 structural *genes;
2. They are Lac Z* which codes for beta galactosidase*;
3. Lac Y* which codes for lactose permease*;
4. and Lac A* which codes for lactose transacetylase*;
5. structural genes have a common promoter* sequence upstream which has a RNA
polymerase binding site.
6. Situated within the promoter is also a Catabolic activator protein binding site*.
7. The operator* (situated between the promoter and the structural genes) has a site
which allows for an activated repressor to bind.
8. The is a terminator sequence downstream at the end of the lac operon;
9. Further upstream of the operon is the regulator gene Lac I which codes for the repressor
protein;

9(b) Explain how the presence of lactose in the growth medium switches on the lac operon. [8]

1. In the absence of lactose, a basal level of -galactosidase and permease is present

within the cell because repression of the lac operon by the repressor is ‘leaky’
(Weak interactions between the repressor and operator means that the repressor may
dissociate from the operator occasionally to allow the RNA polymerase access to the lac
operon)
2. A small number of permeases* is therefore present to transport lactose from the
surrounding medium into the cell;
3. Some lactose will be converted to its isomer allolactose* by -galactosidase* in the
cell;
4. Allolactose then acts as an inducer* molecule which binds to the repressor protein at its
allosteric site;
5. This alters the conformation of the DNA-binding site of the repressor
6. Such that the repressor is inactivated and is no longer complementary in shape and
charge to the operator and thus cannot bind to the operator;
7. RNA polymerase* is now free to bind to the promoter* and can move downstream to
transcribe* the structural genes to form a polycistronic mRNA;
8. Glucose is absent thus high level of cAMP will activate the CAP which binds to the
promoter of the lac operon to increase the frequency of transcription.
9. operon is now turned on with increased synthesis of -galactosidase*, permease* and
transacetylase* for metabolism of lactose (positive feedback)

9(c) Suggest why operons are necessary in bacteria. [4]

1. Clustering genes of related functions together under the control of a promoter allows for
regulation of gene expression*;
2. This allows bacteria to only produce enzymes* when required and allow quick
responses to changes in the external environment;
3. This allows efficient use of energy and conservation of resources/prevent waste of
energy;
4. This ability to adapt to changes in the environments confers a selective advantage* to
the bacteria;
5. This also allows bacteria to be able to use a variety of sugars / substrates;
11
Raffles Institution Nov 2007 (H2 Biology) Paper 2 (for 9744 syllabus)

Nov 2007 H2 Bio Paper 3

N07P3Q1 (OUT OF SYLLABUS)

N07P3Q2
Fig 2.1 shows the three stages involved in the polymerase chain reaction (PCR).
a) With reference to Fig.2.1, explain what is happening at each of the three stages. [2m
each stage]
Stage 1
1. heating to 95 C separates two strands of the DNA double helix;
2. by breaking the hydrogen bond between complementary bases through increased
molecular vibrations, denaturing the DNA;

Stage 2
3. Cool to around 64 C, for DNA primers to anneal;
4. Primers anneal to complementary 3’ end of each template/single strand;

Stage 3
5. At 72 C , Taq polymerase* synthesizes complementary DNA strand;
6. Chain extension occurs from3’ OH end of primer;

Teachers’ comments:
Appropriate temperatures were not suggested. Point 2 and 6 often left out. The importance of
primer providing a free 3’ OH was often overlooked. For point 4, primers anneal to the flanking
regions of the target DNA sequence, not to the ends of the DNA fragment. The target DNA
fragment can be huge!

Fig. 2.2 shows what happens if the PCR is repeated.

b) With reference to Fig.2.2,
(i) State how many copies of the original DNA there will be after 25 cycles, [1] \

226* (not 225 since 1st cycle is 22 and 2nd cycle is 23 etc)

(ii) Explain how PCR facilitates genetic fingerprinting. [2]

1. PCR amplifies DNA from a limited source of DNA especially important in a crime
scene where tissue evidence is limited;
2. So that there is sufficient amount of DNA for procedures involved in analysis as the
bands may not be visible if DNA is limited;

Teacher’s comments:
Many students simply stated the obvious feature of PCR – amplifying DNA without explaining
why you needed so much DNA. Even though students mentioned that it generates sufficient
amount of DNA to be analysed, I cannot tell for certain that they know that visualizing the bands
require sufficient DNA. The fingerprint isn’t the source of the DNA! It is named as such because
it is similar to the use of a traditional fingerprint in identification.

1
Raffles Institution Nov 2007 (H2 Biology) Paper 2 (for 9744 syllabus)

c) Legislation in many countries prohibits the import and export of endangered species.
Explain the process of genetic fingerprinting and suggest how it can be used to verify
whether an organism belonging to an endangered species has been bred in captivity. [5]

1. In genetic fingerprinting, genomic DNA is extracted and;

2. digested with a restriction enzyme;
3. Due to polymorphic nature of DNA in different individuals, there will be variations in
number and location of restriction sites and number of tandemly repeated nucleotide
sequences (e.g. micro and minisatellite loci) among individuals
or convey idea of RFLPs;
4. Result in a unique* banding pattern among individuals;
5. After separation by gel electrophoresis, the DNA fragments are transferred to
nitrocellulose membrane;
6. Incubate with radioactive probe(s) + expose on x-ray film/autoradiography;
7. Genetic fingerprint of suspected organism can be compared against fingerprint of
captive organisms and wild organisms to see how closely related they are;
8. If fingerprint pattern is similar to pattern of captive organisms, then they have been
bred in captivity;
Note: Two marks must come from pts 7 and 8. (Verifying whether an organism belongs
to a species bred in captivity)

Teacher’s comments:
Generally ok but some students didn’t answer the question of how to verify the origins of the
said animal. Students should go on to illustrate how a conclusion can be made if a certain
pattern emerges. Many simply said compare said animal with captive animals and left it open
ended.(pt 8 missing) Some simply mentioned that Southern blot was performed without the
details. Few had point 3. [Total: 14]

N07P3Q3(OUT OF SYLLABUS)

N07P3Q4(OUT OF SYLLABUS)

[Total: 20]

2
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

Nov 2008 H2 Bio Paper 2

N08P2Q1

(a) With reference to Fig. 1.1,

(i) identify the two main stages of mitosis when these movements occur,[2]
1. Metaphase* from 0 to 15mins (Quote from graph);
2. Anaphase* from 15 to 30mins

(ii) describe what happens to the chromatids after 15 minutes,[2]

1. After 15min, the centromeres divide/separate* (R: split, replicate) and
2. sister chromatids* separate and each sister chromatid is now a daughter
chromosome*
3. Kinetochore microtubules* shorten & pull the chromosomes to opposite
poles*. (R: ends) after 30 min the distance between the centromeres increases to
48 µm; (quote values)

(iii) account for the changes in curve C.[2]

1. For the first 10minutes, the kinetochore microtubules* shorten and the
centromeres do not divide. This pulls the poles* closer causing the initial
decreases in distance between poles; 45µm to 40 µm;

2. After 25 minutes distance between pole increases from 45um to 50um as they
move apart. This is because the non-kinetochore microtubules* elongate and
slide pass each other causing the cell to elongate to prepare for cytokinesis

(b) Outline the role of centromeres.[3]

1. They are non-coding tandem repeat sequences at one location along the length
of a chromosome where sister chromatids* to adhere to each other and allow
proteins called kinetochores*, and subsequently kinetochore
microtubules/spindle fibres*, to attach;
2. Thus allowing the alignment of chromosomes at the equator* during
metaphase* and
3. Subsequent separation of sister chromatids, the chromosomes formed can be
pulled to opposite poles*; R ends
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

(c) State four ways in which the behaviour of chromosomes in meiosis differs from their
behaviour in mitosis.[4]
Pt of comparison Mitosis Meiosis

Pairing of 1 Homologous chromosomes* 1 Homologous

homologous do not pair up via synapsis* chromosomes* pair up by
chromosomes and there is no formation of a process called
bivalent* synapsis* to form a
bivalents* at prophase I*.

Crossing over 2 There is no chiasma formation

and crossing over and hence no 2 Chiasmata* formation and
exchange of equivalent portion crossing over* occurs
of genetic material or alleles such that exchange of
occurring between homologous equivalent portion of
chromosomes. genetic material or alleles
occur between non-sister
chromatids* of
homologous
Arrangement of 3 Chromosomes arranged singly chromosomes* during
chromosomes at at the metaphase prophase I*
Metaphase plate/equator* during
metaphase. 3 Homologous
chromosomes arranged as
pairs at metaphase
Behaviour of 4 It involves division of plate/equator* during
chromosomes at centromeres and separation of metaphase I*
Anaphase sister chromatids* at
anaphase*.
4 It involves the separation
of homologous
chromosomes* at
anaphase I* .
Centromeres do not
divide at anaphase 1. and
division of centromeres
and separation of sister
chromatids* at anaphase
II*

[Total: 13]
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

N08P2Q2

(a) State the type of mutation responsible for this change in the amino acid sequence. [1]
A single base substitution*.

(b) Explain the significance of the change in amino acid to the properties of haemoglobin.
[3]
1. Hydrophilic* charged glutamic acid* is replaced by hydrophobic*, non-polar
valine*;
2. At low O2 concentrations, loss of O2 from HbS* results in an unusual conformational
change that causes a hydrophobic patch to stick out;
3. This hydrophobic patch attaches to a hydrophobic patch on another HbS causing
them to polymerise* into insoluble fibers*;

(c) Describe the effects of this change in the haemoglobin.[3]

1 Long insoluble HbS fibers* within red blood cell* causes its shape to be distorted
from a normal biconcave shape to a sickle* shape;
2 Sickle red blood cells are more fragile resulting in them having a shorter life span
results in shortage of red blood cells and poor oxygen transport resulting in
anaemia;
3 Sickle-shaped red blood cells, being pointed and elongated, may also get lodged in
small blood vessels (capillaries) and therefore interfere with blood circulation. This
may result in organ damage;
(note: haemoglobin is not = to red blood cells!)

(d) (i) Suggest how formation of HbF would be induced.[2]

1. Hydroxyurea has ability to activate transcription* of HbF gene in adult resulting
in HbF mRNA*;
2. HbF mRNA is in turn translated* to form HbF;

(ii) Suggest how elevated levels of HbF may reduce the symptoms of sickle cell
anaemia.[2]
1. Elevated levels of HbF will result in a relatively lower number/proportion of HbS*
in red blood cell. Less polymerisation of haemoglobin occurs,;
2. This leads to fewer sickled cells and less cell lysis, resulting in a longer life span
of red blood cell and hence improved oxygen carrying capacity of the blood;
3. Compared to HbS, HbF has a greater affinity for oxygen;

[Total: 11]

N08P2Q3

(a) Describe how an enveloped virus such as H5N1 virus enters a host cell.[3]
1. Haemagglutinin* binds to sialic acid receptor* on host cell membrane
2. Virus then enters host cell by endocytosis*: where host cell membrane
invaginates* and pinches off, placing virus in an endocytotic vesicle*.
3. Vesicle then fuses* with a lysosome*, and the resulting drop in pH stimulates
fusion of viral envelope with vesicle membrane
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

4. Releasing nucleocapsid* into cytosol

(b) State how, despite having no ribosomes, the influenza virus is able to trigger the
synthesis of these proteins.[1]
It makes use of the host cell’s ribosomes* for protein synthesis.

(c) The RNA segments containing genes PB2, PBt and PA code for the RNA
polymerases shown in Fig. 3.1. Explain why such RNA segments are needed by the
virus.[2]
(note: Influenza virus uses RNA-dependent RNA polymerase)

1. Such RNA segments code for RNA-dependent RNA polymerases which are not
present in the host cell.
2. The RNA-dependent RNA polymerases are required for the replication of viral
RNA,
3. And synthesis of viral mRNA used for translation into viral proteins.

(d) Explain how inhibiting neuraminidase can prevent influenza.[2]

1. When neuraminidase is inhibited, it cannot cleave the sialic acid to release the
viruses from the infected cells
2. to infect other cells
(Note: Tamiflu, aka Oseltamivir, is a competitive inhibitor for neuraminidase,
structurally similar to sialic acid)

[Total: 8]

N08P2Q4

4 (a) Explain how one named factor can increase the chances of a cancerous growth.[3]
1. exposure to ultraviolet light / radioactivity / ionizing radiations, (mention of
radiation alone not sufficient), Carcinogens such as tar in cigarette smoke,
asbestos, benzene, formaldehyde, ethidium bromide, Viruses like avian sarcoma
virus, HPV;
2. viruses like avian sarcoma virus, HPV, excessive UV light / radioactivity /
carcinogens like asbestos, benzene, formaldehyde (any carcinogen);
3. increase chances of DNA damage and mutations (e.g. of mutation);
4. leading to loss of function mutation of tumour suppressor genes and / or gain of
function mutation of proto-oncogenes to form oncognene;
5. disruption of these regulatory genes will lead to excessive cell proliferation and
inability to repair damaged DNA;
6. resulting in formation of a maglinant tumour capable of metastasizing to other
parts of body to form secondary tumours;

(b) State what you understand by the terms,

(i) proto-oncogene [2]
1. found in normal cells;
2. gene that codes for a protein;
3. involved in normal cell proliferation/division;

(ii) oncogene [2]

1. mutated form of proto-oncogene* such that
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

2. there is an excessive production of the protein or oncogene codes for a protein

with increased activity / more resistant to degradation, leading to;
3. greater than normal / uncontrolled cell proliferation/division resulting in cancer

(c) Explain how the Myc proto-oncogene becomes an oncogene in Burkitt's lymphoma.[3]
1. Idea of Proto-oncogenes are normal genes found in normal cells: Myc gene in
chromosome 8 is regulated by a normal promoter / expressing normal amount of
Myc protein;
2. Mechanism how proto-oncogene becomes oncogene: Translocation is a
chromosomal mutation when Myc gene being transferred from chromosome 8 to
chromosome 14 to be under the influence of an enhancer* of lgH gene;
3. Enhancement of gene expression: resulting in high level expression/ transcription
of Myc gene to produce excessive amount of Myc protein/excessive signal
molecules or growth factors;
4. thus this is a gain-in-function mutation;
5. Excessive amount of Myc protein stimulates cells to undergo greater cell division ,
leading to tumour formation;

(d) Suggest how amplification of Mdr1 will allow tumour cells to show multi-drug
resistance[2]
1. Gene amplification resulting in more copies of Mdr1 gene, therefore more
membrane-bound transporters transcribed and translated to ensure;
2. greater pumping of drugs out of cells so that drugs cannot accumulate and reach
toxic levels for cells greater resistance;

(e) Suggest how this multi-drug resistance in a tumour may be overcome.[2]

1. anti-sense RNA that will bind to mRNA transcripts of Mdr1 gene;
2. to prevent translation of membrane-bound transporter so that number of
transporters will decrease;
OR
1. molecular inhibitors that bind directly to membrane-bound transporters;
2. to prevent their pumping activities so that there will be no efflux of the anticancer
drugs;

[Total: 14]
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

N08P2Q5

(a) Draw a genetic diagram in the space below to show the cross described.[5]

Parental phenotype Purple grain X Purple grain

Parental genotype RrEe RrEe

Meiosis
Gametes (n) RE Re rE re X RE Re rE re

Random fertilisation
Male gametes

RE Re rE re

RREE RREe RrEE RrEe

RE Purple Purple Purple Purple
Female gametes

RREe RRee RrEe Rree

Re Purple Red Purple Red

RrEE RrEe rrEE rrEe

rE Purple Purple white white

RrEe Rree rrEe rree

re Purple Red white white

Offspring genotypic ratio 9 R_E_ : 3 R_ee : 3rrE_ : 1rree

4
Offspring phenotypic ratio 9 purple : 3 red : 4 white
[5]
(b) Explain why it would be useful to carry out a chi-squared test on these results. No
calculations are required to answer this question.[2]
1. To determine if the difference between observed phenotypic ratios (in this case
9:3:4) and expected 9:3:3:1 dihybrid ratio
2. was significant, or due to chance;

(c) State the name for this type of interaction between gene loci. [1]
1. epistasis

(d) Explain how different genotypes give rise to a white phenotype.[3]

1. rr is epistatic over E and e locus* (Recessive epistasis);
2. rr prevents the formation of any pigment hence rrEE, rrEe or rree will give only white
phenotypes;
3. R_ produces enzyme R which converts a colourless precursor to red
4. And with presence of E_, the red pigment is converted to a purple pigment
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

(e) Explain how you could determine that a plant grown from a white grain was
homozygous recessive at both loci.[3]
1. Cross this plant with unknown genotype with a pure breeding red plant (RRee);
2. Resultant offspring would have either all red or red & purple grains;
3. If purple grains seen, then the original plant must have at least one E allele at the
E/e locus;

N08P2Q6

Fig. 6.1 shows the sequence of events in the Jak-STAT cell signalling pathway.
(a) Describe the stages A and B shown in Fig. 6.1.
A [2]
Stage A:
1. Binding of α-interferon (ligand) to receptor tyrosine kinase (RTK), α-interferon
receptor;
2. causes the two receptor subunits to aggregate/dimerise*;
3. This is followed by cross-phosphorylation* of Tyk2 and Jak1 by intrinsic kinase
activity of receptor;

(Tyr2 and Jak1 are different forms of the Janus kinase)

Note: Please use cross-phosphorylation NOT auto-phosphorylation as indicated by
the arrows in the diagram.

Stage B
4. Activated Tyk2 and Jak1 will in turn phosphorylate tyrosine residues* on
intracellular domain* of the α-interferon receptor;
5. STAT1 and STAT2 proteins will bind to the activated tyrosine residues via SH2
domains and will in turn become phosphorylated;
6. STAT1 and STAT2 then dissociate from α-interferon receptor and the two subunits
dimerise;

(b) Explain why interferon cannot act directly on the DNA in the nucleus.
1. Interferon is too large, cannot pass through any transient pores form within cell
surface membrane;
OR
2. cannot pass through hydrophobic core of cell surface membrane because it is
polar and will be repelled;

(c) Jak-STAT and other cell signalling pathways are advantageous for multicellular
organisms. Describe three advantages of such a cell signalling pathway.[3]

1. Facilitate signal amplification as only a small number of signal molecules (ligands) is

needed to solicit a large response from cell;
2. Provides multiple checkpoints for regulation so that the cellular response can be
regulated and controlled at each step;
3. Multiple responses to 1 signal molecule (ligand) because 1 signal molecule (ligand)
can trigger multiple signal transduction pathways to elicit different responses;
4. Ensures specificity because the specific signal molecule (ligand) binds to a specific
receptor (ref. to specific conformation) to elicit specific reaction via specific pathway
in each cell type;
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

5. Ability of a signal molecule (ligand) to activate genes in nucleus upon binding to cell
surface receptor without the need to move into nucleus;
6. Ability of a signal molecule (ligand) to activate many different cells simultaneously;
[Total: 9]

N08P2Q7

7 (a) Describe the environmental factors that act as forces of natural selection on finches
in the Galapagos Islands.[3]
1. Different types of food available in the environment / habitat e.g. seeds, cactus
flowers, cactus fruits;
2. If the seeds available in the environment / habitat are hard or large, finches with low
beak depth will not be able to crush these seeds and hence selected against;
(Accept elaboration of beak phenotypes that are selected for)
3. Limited availability of food results in competition which exerts a selection pressure to
exploit varied food sources;

(b) Describe, using information from Fig. 7.1, how these two genes interact to produce
the range of beak shapes.[2]
1. Increase in BMP4 expression increases beak height/depth and width;
2. Increase in CaM expression increase beak length;
3. Moderate range of BMP4 expression level with high or low CaM expression give
rise to intermediate beak depth/width;

(c) Explain how the molecular evidence described supports Darwin's theory of natural
selection. [4]
1. BMP4 gene and CaM gene are homologous genes present in the common ancestor
and all its descendants;
2. There was modification in terms of different levels of gene expression in the two
genes resulting in variations in beak shape in the population; (students tend to miss
this out)
3. Selection pressure* is the type and availability of food;
4. Finches with beaks suited to feed on a particular type of food available in the
environment are selected for and will survive;
5. and reproduce fertile, viable offspring to pass on their favorable alleles to their next
generations; (students tend to miss this out)

(d) Suggest the role of the islands in the evolution of so many species of finch. [2]
1. Islands provide means of geographical isolation* by being surrounded by bodies
of water;
2. such that gene flow is disrupted* between islands that may cause speciation;
3. Different islands have different environments thus exposing the finches to different
selection pressures* and the finches well adapted for that environment will be
selected for and survive to produce fertile, viable offspring;
[Total: 11]
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

N08P2Q8

(a) Describe how the Calvin cycle differs from the Krebs cycle. [7]
Krebs cycle Calvin cycle
Site Occurs in the matrix in the Occurs in the stroma in the
mitochondrion. chloroplast
+
Electron/hydrogen NAD /FAD accepts the Reduced NADPH is oxidized
carrier(s) electrons and proton / reduced to become NADP+.
to form NADH/FADH
NAD and FAD NAPDH
Carbon dioxide Produced Incorporated into sugars
ATP Produced during substrate Used up
level phosphorylation
Nature of process Catabolic process, releasing Anabolic process, converting
energy stored in acetyl CoA to energy in the form of ATP
ATP and reduced NADH, and NADPH to producing
FADH. starch.

(b) Explain the small yield of ATP under anaerobic conditions in both yeast and
mammals. [8]
1. In the absence of oxygen, the final election acceptor*, members of the
electron transport chain that have previously picked up electrons and protons
from NADH/FADH2 cannot be re-oxidised;
2. oxidative phosphorylation* eventually stops as no further electrons are passed
down the chain, hence no ATP will be produced via chemiosmosis.
3. NAD and FAD cannot be regenerated / NADH and FADH2 will remain reduced,
hence Krebs cycle and Link reaction cannot proceed.
4. No ATP produced via Krebs cycle via substrate level phosphorylation*.
5. The regeneration of NAD* in cytoplasm allowed glycolysis to proceed,
6. produced 2 net ATP per glucose molecule via substrate level phosphorylation*.
7. In yeast cytoplasm, pyruvate is reduced to lactate by NADH Or pyruvate is first
decarboxylated to ethanal, then reduced to ethanol by NADH via alcoholic
fermentation;
8. in mammals cytoplam, pyruvate was reduced by NADH to lactate via lactate
fermentation.

(c) State the similarities between ATP production in mitochondria and chloroplasts and
suggest why these similarities exist. [5]
Similarities
ATP synthesis is both organelles occur through chemiosmosis*;, where in both ,
e- are passed down a series of electron carriers that are progressively more
electronegative;
energy is released and it is coupled to the pumping of protons/H+, (from the
mitochondrial matrix into the intermembrane space in mitochondria, and from
stroma to thylakoid in chloroplasts.)
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

results in a build up a proton-motive force / proton gradient* across the inner

mitochondrial membrane and thylakoid membrane.
H+ flows down its concentration gradient through ATP synthase*,
Energy from Proton motive force used to phosphorylated ADP to form ATP..
These similarities exists as mitochondria and chloroplast are thought to be originated as
separate prokaryotic organisms that were taken inside a bigger prokaryotic and be part
of it (ref to endosymbiotic theory).

N08P2Q9

a) Describe how classification differs from phylogeny. [7] (OUT OF SYLLABUS)

Basis of Classification Phylogeny

comparison
1 (Basis of) Classification groups Whereas phylogeny traces the
grouping of organisms based on evolutionary history of organisms
organisms overall/morphological based on ancestor-descendent
similarities and does not take relationships;
into account the evolutionary
history of organisms
2 System of It is a system of naming. It Organisms are arranged based
organizing involves grouping organisms on their evolutionary relationship
organisms into a kingdom, phylum, with each other with each
class, order, family, genus organism assigned a position on
and species using a a branching tree relative to other
hierarchical classification* organisms;
system
3 How species Uses binomial Uses a phylogenetic tree*
are presented nomenclature* where related organisms are
grouped into the same branche
on a phylogenetic tree;
4 Nature of Being based on Makes use of homologous
characteristics morphological characters, characters* that are derived
does not discriminate from a common ancestor to
between analogous or group organisms;
homologous characters
5 Types of Uses only morphological Uses morphological anatomical,
characters characteristics embryological as well as
used molecular characteristics such
as DNA, RNA, amino acid
sequences; Also fossil records.
6 Strengths and Easily place an organism into Cannot immediately place an
weaknesses its well defined group. organism into the phylogram as
evolutionary history need to be
established from multiple
May wrongly classify sources;
organisms that are not Rarely classifies wrongly as
related but look similar due to convergent evolution will be
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

convergent evolution* placed in separate branches;

7 Inference of Does not allow inference of While phylogeny indicates
speciation historical speciation events speciation events as the node of
events the phylogenetic tree;
8 Inference of Cannot infer of how closely Allows accurate inference of
relationships related 2 species are how closely related 2 species
especially since they are are by looking at how recently
grouped together in the same they diverged from their common
hierarchy, “species” ancestor;
9 Inference of Does not allow inference of Allows inference of common
common common ancestors ancestors. Descendants of a
ancestors common ancestor are
represented in the same branch;
10 Application of Not possible to apply If molecular evidence is used,
molecular molecular clock to date can apply the molecular clock to
clock speciation events infer time of speciation;

b) Explain the advantages of molecular methods in classifying organisms. [8]

1. All known life is based on nuclei acids thus studies involving any types of taxa can
use DNA sequence data.
2. They can be used to compare species that are morphologically indistinguishable
and to assess relationships between groups of organism that are so
phylogenetically distant that they share very few morphological similarities.
3. They are objective* and quantitative*. Molecular character states are
unambiguous (e.g. A, C, G and T) whereas some morphological characters, such
as those based on the shape of a structure, can be less easy to distinguish
because of overlaps between different character states
4. Amino acid sequences for many proteins and nucleotide sequences for a rapidly
increasing number of genomes can be accessed from electronic databases and
used for comparative study and classification.
5. Offers a large and essentially limitless set of characters to be studied. Each
nucleotide position can be considered a character and organism has millions -
billions of nucleotide positions
6. A single experiment can provide information on many different characters: in a DNA
sequence, for example, every nucleotide position is a character with four character
states A, C, G and T. Large molecular datasets can therefore be generated
relatively quickly.
7. Molecular data are easily converted to numerical form and hence are amenable to
mathematical and statistical analysis.
8. The amino acid sequences of protein and nucleotide sequences of RNA, DNA
molecules provide the most detailed, and unambiguous data for molecular
phylogenetics.
9. The degree of relatedness can be inferred and quantified by calculating the number
of nucleotide differences between species
10. Molecular methods provide more characters than morphological characters to
classify organisms as molecular differences are not visible phenotypically,
especially if introns and intergenic spacers are compared
11. Molecular characteristics can clearly show if organisms are wrongly classified due
to morphological characters that are similar between organisms due to convergent
evolution
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

c) Suggest how viruses evolved. [5]

[guide: the question is asking you to suggest the origins of viruses more than its
evolution through natural selection] http://www.nature.com/scitable/topicpage/the-
origins-of-viruses-14398218

Viruses did not come from a common ancestor

1. Viruses are very diverse, having RNA or DNA genomes, being double stranded or
single stranded, having circular or linear nucleic acid molecules, contain one or
many nucleic acid molecules pointing to a very different evolutionary
origin/paraphyletic nature for each one of them;
2. Their replication strategies are also very diverse, some using reverse transcriptase
while others use RNA dependent RNA polymerases and yet others requiring DNA
polymerase for its replication;
3. The polyphyletic nature of the diverse groups of viruses therefore suggest that they
do not share a common ancestor*;

The possible evolutionary origins of virus

4. The progressive hypothesis: They are genetic material that can move within a
genome called retrotransposons. Their life cycle resembles that of HIV and other
retroviruses and they encode reverse transcriptase as well. We can speculate that
they acquired a few structural proteins that would allow them to exit a cell and enter
a new cell;
5. The regressive hypothesis: The existing viruses may have evolved from more
complex, possibly free-living organisms that lost genetic information over time, as
they adopted a parasitic approach to replication. e.g. are large DNA viruses with
large genomes like smallpox and mimivirus;
6. The virus-first hypothesis: The first two hypothesis suggest that cells exists first
before viruses. Here viruses evolved first in a precellular world. The very first
replicating molecules are RNA, not DNA. We know that some RNA molecules,
ribozymes exhibit enzymatic properties. Viruses could be the first replicating
entities. Eventually enzymes for the synthesis of membranes and cell walls evolved,
resulting in the formation of cells;

Viruses can still change/evolve (not the focus)

7. They are all parasites that multiply only within a host cell and it is also within the
host that they can accumulate mutations and change/evolve through a process
called antigenic drift;
8. Or viruses went through antigenic shift where 2 different viruses may have infected
a single host cell and recombined into a new virus. E.g H7N9 a recombination of
viruses from wild migratory birds and chicken avian influenza virus with ducks as a
host;
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

Nov 2008 H2 Bio Paper 3

N08P3Q1

Q1 (a) Explain,

(i) what is meant by a restriction enzyme [3] (OUT OF SYLLABUS)

1. Enzyme that recognizes and bind a specific 4-6 base pair DNA sequence called
a restriction site as its active site is complementary to the DNA sequence; Note:
Binding must be mentioned.
2. Enzyme cuts/breaks phosphodiester bonds on specific positions on both DNA
strands;
3. Giving rise to either blunt or sticky ends depending on type of restriction enzyme;
4. Used as a defense mechanism by bacteria against bacteriophage by cutting up
foreign DNA, hence restricts multiplication of viruses;

(ii) how gel electrophoresis is used to separate fragments of DNA.[5]

1. The fragments of DNA are pipetted into the wells at the top of the gel in a position
furthest from the positive electrode/anode;
2. Negatively-charged DNA;
3. Migrates out of well towards direction of +ve electrode/anode when subjected to
an electric field / current;
4. Fragments migrate through agarose gel matrix, made up of a meshwork of
polysaccharides;
5. Meshwork impedes movement of longer fragments more than shorter fragments;
6. Longer fragments migrate slower compared to shorter fragments;

(b) With reference to Fig. 1.1, calculate the percentage increase in length of DNA
fragment F when the mutant allele is cleaved by Mstll, compared with fragment F from
the normal allele. Show your working.
1. answer ..........................18.% [2]
2. 0.2/1.1 x 100 = 18.2%

(c) (i) On Fig. 1.2 identify DNA fragment F of the mutant allele by means of a labelled
arrow. Put your answer on Fig. 1.2. - the 1.3kb fragment (Indiv B) [1]

(ii) With reference to Fig. 1.2, explain the banding pattern of individual C.[4]
1. C is heterozygous for sickle cell;
2. Normal and mutant allele are on same gene loci on a pair of homologous
chromosomes;
3. Normal allele gives rise to an intermediate and short fragment upon restriction
digestion with MstII and these fragments correspond to the middle and bottommost
bands respectively;
4. Mutant allele gives rise to a single long fragment upon restriction digestion with
MstII and this corresponds to the uppermost band;

1
Raffles Institution Nov 2008 (H2 Biology) Paper 2 (for 9744 syllabus)

N08P3Q2

Q2 (a) (i) Explain what is meant by a stem cell.[2]

1. A stem cell is an undifferentiated/unspecialized* cell capable of undergoing
extensive proliferation* and self-renewal*;
2. and retains the potential to differentiate to produce specialized cells upon receiving
appropriate molecular signals;

(ii) With reference to one example, outline the normal roles of stem cells in living
organisms.[2]
1. Haematopoietic stem cells from the bone marrow;
2. divide and differentiate giving rise to blood cells including, red blood cells, white
blood cells to replace those that have been worn out;
Any other valid example; with role; e.g. zygotic stem cells, embryonic stem cells

(b) With reference to Fig. 2.1,

(i) describe two different types of mutation of the gene coding for gamma-c that could
each result in no gamma-c being formed [2]
1. A substitution* mutation resulting from one base being replaced by another may
result in a different amino acid being incorporated into the polypeptide, leading to
different protein being formed/a premature stop codon*, leading to truncated
polypeptide chain;
2. A deletion* mutation where a nucleotide (or more) was removed from the gene
resulting in a frameshift that will cause all the amino acids incorporated after the
point of mutation to be incorrect /introduce a premature stop codon, leading to
truncated polypeptide chain;

Any other mutation that leads to non-functional protein product with explanation of
the mutation;

(ii) describe one non-viral gene delivery system that could have been used to insert the
normal allele of the gamma-c gene into stem cells [3] (OUT OF SYLLABUS)

(iii) suggest the advantages of using a viral gene delivery system.[3](OUT OF

SYLLABUS)

(c) Suggest why the more recent gene therapy has proved more effective.[3]
1. As stem cells are able to proliferate* and self-renew*, there is a steady supply of
stem cells that can continue to generate more cells so treatment is
permanent/long term;
2. Genetically modified stem cells can differentiate* into lymphocytes that also
carry the normal functional allele and so function normally;
3. If lymphocytes were genetically modified, the therapeutic effect wears off after the
modified lymphocytes die and are not continually replaced; (??any other ideas)
[Total: 15]
N08P3Q3(OUT OF SYLLABUS)

N08P3Q4(OUT OF SYLLABUS)

2
Raffles Institution Nov 2009 (H2 Biology) Paper 2 (for 9744 syllabus)

Nov 2009 H2 Bio Paper 2

N09P2Q1
Fig. 1.1 shows ribosomes attached to the endoplasmic reticulum, carrying out protein
synthesis.

a) Name the structures labeled A to C.[3] (3 lines)

A : mRNA;
B: protein/polypeptide being synthesized;
C : cisternal space of rough endoplasmic reticulum;
1 mark each

b) Ribosomes are found in several locations in the eukaryotic cell. Place a tick, (√) in the
correct column to show these locations. [2]

location ribosome present ribosome absent

nucleus √
chloroplast √
vacuole √
mitochondria √

c) Describe the function of the protein that forms the membrane channel. [3] (5 lines)
1. Allows transport of large polypeptides into the cisternal space/lumen as these cannot
traverse the hydrophobic core of the phospholipid bilayer of the Rough ER;
2. Channel protein functions as a receptor to bind (to the signal recognition particle which in
turn binds) to the signal peptide sequence of the polypeptide;
3. Channel protein also holds the bound ribosomes in position on the Rough ER.

Note: The channel protein on the RER membrane is called the translocon.

d) Proteins synthesized by ribosomes attached to the endoplasmic reticulum may be

transported out of the cell. Outline the route taken by such a protein.[4] (7 lines)
1. Proteins enter the lumen of the cisterna of the rough endoplasmic reticulum;
2. Transport vesicles bud off from the Rough ER and carry the proteins to the Golgi
apparatus. They fuse with the ‘cis’ face of the Golgi apparatus
3. A secretory vesicle containing the modified protein will then bud off from the ‘trans’ face
of the Golgi apparatus;
4. and fuse with the cell surface membrane, releasing the protein content of the vesicle by
exocytosis;
5. Microtubules direct the movement of the transport vesicle to the Golgi apparatus and the
secretory vesicle to the cell surface membrane.

e) Suggest how prokaryotes, which have no endoplasmic reticulum or vesicles, are able to
secrete proteins.[1] (2 lines)
They secrete proteins via protein channels/pores on their cell surface membrane.
Raffles Institution Nov 2009 (H2 Biology) Paper 2 (for 9744 syllabus)

N09P2Q2

a) State two ways in which a plasmid such as the F plasmid differs from the bacterial
chromosomes. [2]
1. F plasmid has significantly fewer base pairs/ is much smaller than bacterial chromosome
2. F plasmid contains non-essential genes (not required for cell survival)/genes which
confer advantages such as antibiotic resistance, unlike bacterial chromosome which
codes for essential genes such as enzymes for cell metabolism

b) Outline the processes of

(i) bacterial conjugation [2] (3 lines)
1. Sex pilus* of F+ bacterial cell makes contact with a F- cell and retracts to bring the F-
cell closer so a mating bridge* is formed between the 2 cells
2. One of the 2 strands of the plasmid DNA in F+ cell is nicked and transferred from the
F+ cell to the F- cell through mating bridge (via rolling circle mechanism) as the other
DNA strand is used as a template for elongation
3. The single strand F plasmid DNA circularizes in F- cell and is used as a template* to
synthesise a complementary strand for a double-stranded F plasmid DNA resulting in
F+ cell
Note: Details of rolling circle (RC) mechanism probably not required. RC is used when
discussing DNA replication and not the transfer. In this case, these 2 are concurrent events.
Also, this question is only 2 marks.

(i) bacterial transformation. [2] (3 lines)

1. Fragments of foreign naked DNA (from lysed bacterial cells) in the surrounding
medium are taken up by a bacterial cell via surface proteins.
2. The foreign DNA is incorporated into the bacterial chromosome/DNA via crossing
over/homologous recombination*
3. If the foreign DNA contains a different allele that is now expressed in the bacterial
cell, the bacterial cell has transformed.

(c) Suggest two potential benefits of conjugation for the recipient bacteria. [2]
1. Gains new alleles that, when expressed, allow it to survive in a different environment.
E.g. antibiotic resistance,
2. use of a new metabolites/resources e.g.new carbon source by producing the relevant
enzyme/s
(can receive relatively large amounts of genetic material/DNA at once)

(d) Outline the main stages in bacterial transduction. [3] (5 lines)

Describe either specialized on general transduction.
General transduction
1. A phage* infects a bacterium*, injecting its viral genome into the host cell
2. The bacterial DNA is degraded into small fragments, one of which may be randomly
packaged into a capsid* head during the spontaneous assembly*of new viruses
3. Upon cell lysis, the defective phage will infect another bacterium and inject bacterial DNA
from the previous host cell into the new bacterium
4. The foreign bacterial DNA can replace the homologous region in the recipient cell’s
chromosome if crossing over/homologous recombination* takes place, possibly
allowing the expression of a different allele from the previous host.
Raffles Institution Nov 2009 (H2 Biology) Paper 2 (for 9744 syllabus)

Specialised transduction
1. A temperate phage* infects a bacterium*, injecting its viral genome into the host cell
2. The viral DNA is integrated into bacterial chromosome forming a prophage* which may
be improperly excised to include adjacent segment of bacterial DNA during an
induction* event
3. Bacterial DNA may be packaged into a capsid head during the spontaneous
assembly*of new viruses
4. Upon cell lysis, the defective phage will infect another bacterium and inject bacterial DNA
from the previous host cell into the new bacterium
5. The foreign bacterial DNA can replace the homologous region in the recipient cell’s
chromosome if crossing over/homologous recombination* take place, possibly
allowing the expression of a different allele from the previous host.

N09P2Q3

a) Outline the role of telomeres. [4] (7 lines)

1. Telomeres are non-coding* tandem repeat sequences found at both ends of linear
chromosomes;
2. (non-coding must be mentioned somewhere in answer)
Role – main
3. Each round of DNA replication will result in the shortening of daughter molecules at the
telomeres because DNA polymerase is unable to replace the RNA primers with DNA;
(idea of end replication problem)
4. Since telomeres are non-coding, this ensures that vital genetic information/genes are
not lost / eroded with each round of replication; wtte
Role – others
5. By forming a loop with 3’ overhang, they protect and stabilise terminal ends of
chromosome, hence preventing fusion of the ends with those of other chromosomes;
6. Either: prevent triggering pathways that lead to cell arrest or cell death, because
exposed 3’ overhang will be perceived as DNA damage/DNA double strand break;
OR:
7. prevent DNA repair machinery from recognising the ends of chromosomes as DNA
breaks/damage, hence preventing apoptosis;
8. Either: The 3’overhang of the telomeres allow their own extension, by providing an
attachment point for the correct positioning of the enzyme telomerase in certain cells,
e.g. germ cells
OR:
9. They possess a 3’ overhang which base pairs with the RNA template on telomerase, so
ensures proper alignment of telomerase and allows extension of telomeric ends in
certain cells e.g. germ cells.

b) With reference to Fig. 3.1

(i) describe the function of the telomerase reverse transcriptase [3] (5 lines)
1. Telomerase has an active site* that is complementary in conformation and
charge* to a specific* telomeric DNA sequence;
2. Using telomerase RNA as a template*, telomerase reverse transcriptase forms a
complementary DNA* sequence through complementary base pairing*;
(whereby adenine base pair with uracil, thymine with adenine, cytosine with
guanine, and guanine with cytosine)
3. Catalyzes the formation of phosphodiester bonds* between
deoxyribonuceotides;
4. It then translocates 6 base pairs in the 5’ 3’ direction of the DNA overhang to
repeat the above processes;
Raffles Institution Nov 2009 (H2 Biology) Paper 2 (for 9744 syllabus)

5. After many rounds, a series of tandem repeats of GGTTAG elongates the

overhang (which can then form a complementary DNA sequence), thus
elongating the telomere/DNA;

(ii) describe the part played by the telomerase RNA. [3] (5 lines)
1. 5 nucleotides of the telomerase RNA anneals and forms complementary base
pairs* with the single-stranded overhang at 3’ end of the telomere;
2. which aligns the telomerase reverse transcriptase wrt the DNA;
3. It serves as the template* for formation of a complementary DNA sequence;
4. whereby adenine pairs with uracil, thymine with adenine, cytosine with guanine,
and guanine with cytosine;
5. Resulting in tandem repeat sequences;

c) Fig. 3.2 shows the base sequence of the telomerase RNA.

(i) The part of the molecule that is directly involved in telomerase activity is within the boxed
area on Fig. 3.2.
Indicate on Fig. 3.2 the precise site that is involved in telomerase activity. [1]

(ii) Suggest why parts of the molecule are double stranded. [2] (3 lines)
1. double stranded nature of parts of telomerase RNA would stabilize the molecule and this
was achieved by complementary base pairing*;
2. It folds into a precise 3D conformation complementary to the active site* of the enzyme;

(iii) Suggest how RNA, such as telomerase RNA, is synthesized. [3] (4 lines)
1. DNA carries the genetic code/gene for the telomerase RNA;
2. RNA polymerase* unzips* the region of DNA containing the gene for the RNA to be
synthesized;
3. Free ribonucleotides then assemble via complementary base pairing* to the DNA
template*;
4. and RNA polymerase* catalyzes the formation of phosphodiester bonds* between
adjacent ribonucleotides;
Raffles Institution Nov 2009 (H2 Biology) Paper 2 (for 9744 syllabus)

N09P2Q4

a) Define the term locus. [2] (3 lines)

1. Locus is the position of a gene on a chromosome
2. Alleles, which are alternative forms of a gene, occupy the same locus
3. Different genes occupy different loci

b) Complete Table 4.1 to show the expected numbers of flies with each phenotype,
assuming there is no linkage. [2]
phenotype observed number (O) expected number if no
linkage (E)
red eye, normal wing 1139 660
purple eye, vestigial 1195 660
wing
red eye, vestigial wing 151 660
purple eye, normal wing 155 660
Total 2640 2640

c) State the meaning of the term linkage in this context. [1] (2 lines)
Linked genes are found on the same chromosome.

d) Draw a genetic diagram to explain the observed results of this test cross. [5]
Use the following symbols,
R red eye; r purple eye; N normal wing; n vestigial wing.

Parental Red eye, Purple eye,

phenotype
Normal wing Vestigial wing

R r r r
Parental X
[1]
genotype
N n n n

R r R r r
gametes [2]

N n n N n

R r r r R r r r [2] for correct

F1 genotype genotypes
linked to
N n n n n n N n
phenotypes
Red eye, Purple eye, Red eye, Purple eye,
F1 phenotypes
Normal wing vestigial wing vestigial wing Normal wing

1139 1195 151 155

Raffles Institution Nov 2009 (H2 Biology) Paper 2 (for 9744 syllabus)

e) Suggest how similar breeding experiments with many different pairs of characters could
be used to map the position of genes on the chromosomes of fruit flies. [3]
(OUT OF SYLLABUS
1. Examine the percentage of recombinants/cross over value, where the percentage of
recombinants/crossover value reflect distance between the pair of traits
Or
Recombinant frequency = total number of recombinant offsprings X 100%
total number of offsprings
2. The greater the percentage of recombinants/crossover value, the greater the distance
between genes (greater chance of crossover)
3. The distance between linked genes is expressed in map units* (or centiMorgan (cM)),
where 1 map unit is equivalent to 1% recombinant frequency
4. If the expected phenotypic ratio is obtained such as 1 : 1 : 1 : 1 when a double
heterozygotes is test crossed, then there is no linkage.

N09P2Q5

a) Suggest why the cytochrome b gene is used to measure changes in DNA sequences in
closely related species. [2]
1. The gene is a homologous gene*, meaning that it is conserved in all the species being
compared and was found in their common ancestor. This forms the basis of comparison;
2. Yet there are sufficient differences/variability in the DNA of cytochrome b for scientists to
distinguish the 3 closely related species;
3. It is found in the mitochondria and hence do not undergo recombination and any
mutation accumulates at a regular rate in the maternal line. Can be used for the
molecular clock;

b) Describe the cause of these changes in DNA sequence. [2] (3 lines)

1. DNA sequence changes when there is a mutation*
2. such as base substitution;
3. The DNA may be damaged by the reactive oxygen radicals generated in the
mitochondrion / mitochondria DNA repair mechanisms may not be as robust;
AVP regarding causes of DNA mutation

c) Describe how these changes in DNA sequences support the neutral theory of molecular
evolution. [3] (5 lines)
1. The graph shows that the changes in DNA sequence occurs in a linear fashion,
suggesting a constant rate of accumulation of mutations over time;
2. This is possible only because the mutations would be selectively neutral and hence has
no selective advantage or disadvantage on its phenotype;
3. Examples of selectively neutral mutations are silent mutations where a change in
nucleotide does not result in a change in amino acid sequence. or conservative
mutations or mutations in non-regulatory, non-coding regions;

d) There are fifty six known (including eighteen extinct) species of Hawaiian honey
creepers. Suggest how this supports Darwin’s theory of natural selection. [2] (3 lines)
1. The large number of species suggests adaptive radiation had occurred;
2. There are different niches in different islands and they each have different selection
pressures present;
3. As a result of variation in populations, favourable genes/traits are selected for via natural
selection in the various niches. These favourable traits/alleles are passed on to the
offspring;
4. Speciation occurs when there is accumulation of mutations and changes in allele
frequencies over time between the sub-populations due to disruption of gene flow;
Raffles Institution Nov 2009 (H2 Biology) Paper 2 (for 9744 syllabus)

N09P2Q6

a) Fig. 6.1 shows the attachment of a chromosome during mitosis.

(i) Name the structures labeled A to C. [3]

A sister chromatids
B centromere
C kinetochore microtubule

(ii) Why does the chromosome appear as a double arm structure? [2] (3 lines)
1. In the S phase of interphase;
2. each chromosome has undergone semiconservative DNA replication to form 2 identical
sister chromatids*;
3. that are held together at the centromere;

b) (i) Name the stage of mitosis shown in Fig. 6.2. [1] (1 line)
Anaphase

(ii) Describe the events that occur during the stage shown in Fig. 6.2. [3] (5 lines)
1. The centromeres divide*;
2. Kinetochore microtubles/spindle fibres which are attached to the centromeres
shorten and;
3. pull the sister chromatids which are now chromosomes to the poles of the cell;

N09P2Q7

a) Explain why the CO2 assimilation rate levels off at high light intensity. [3] (5 lines)
1. At higher light intensity when CO2 assimilation rate levels off, the chloroplasts are light
saturation /saturated* with light;
2. and that photosynthesis is occurring at maximum rate;
3. Light is no longer a limiting factor and other factors such as temperature is the limiting
factor;

b) Explain why the CO2 assimilation rate is negative at the lowest light intensity. [3] (5 lines)
1. At the lowest light intensity there is low/no photosynthesis occurring;
2. because there is less photons of light for photoactivation of chlorophyll molecules in the
light dependent stage of photosynthesis;
3. Respiration occurs at a higher rate than photosynthesis;
Raffles Institution Nov 2009 (H2 Biology) Paper 2 (for 9744 syllabus)

c) Describe the significance of point A to the growth of the plant. [3] (5 lines)
1. Point A is the light compensation point* where at that value of light intensity the rate of
respiration equals the rate of photosynthesis;
2. Amount of CO2 given out during respiration is equivalent to the amount of CO 2 fixed
during the light independent stage of photosynthesis;
3. Thus, there is no net gain in dry mass and no growth as the products of photosynthesis
(e.g. glucose) are used up in respiration;

N09P2Q8

a) Describe the mode of action of enzymes. [8]

Interaction/binding
1. Enzymes have an specific* active site* which is complementary in
shape/conformation and charge* to the substrate;
2. Effective collisions between enzyme and substrate form a temporary enzyme-substrate
complex*;
3. Based on the lock and key* hypothesis, enzyme is the lock* and substrate is the key*
(lock and key hypothesis) Or
4. Based on the induced fit* hypothesis, substrate induces a change in shape in enzyme
active site so that active site is a more precise fit for substrate for effective catalysis.
5. Enzyme-substrate complex held together by weak interactions e.g. hydrogen, ionic
bonds, hydrophobic interactions*

Catalysis
6. Enzyme lowers the activation energy* barrier by: (max of 2 marks)
Aligning substrates next to each other (close proximity) in active site for reaction to
occur
Strain on bonds to be broken / distorts the substrate and reduces activation energy to
achieve transition state
Orientates substrate such that its bonds are exposed to attack
Provide a favorable microenvironment
R-groups of amino acid residues in active site participate in direct catalysis – e.g.
Acid-base catalysis
Release
7. Products no longer fit active site and are released enzyme is unchanged and can be
used again;
Raffles Institution Nov 2009 (H2 Biology) Paper 2 (for 9744 syllabus)

b) Explain how pH affects the rate of an enzyme catalysed reaction. [8]

Rate of reaction
is highest at
optimal pH

symmetrical

1. Draw and annotate the pH response curve of enzyme activity.

2. Each enzyme has an optimum pH at which it is most active. The rate of reaction is
maximum at this optimal pH;
3. Rate decreases as pH deviates from the optimum pH;
4. Excess [H+ ] or [OH- ] ions may affect the ionisation of R groups/side-chains* of amino
acids
5. where excess H+ results in –COO- groups becoming -COOH and excess -OH- results in -
NH3+ becoming –NH2;
6. Hence, disrupt ionic bonds* and hydrogen bonds* that stabilizes the specific*
conformation of active site* denaturation* of enzyme;
7. pH may also change the specific* charge of the R groups* of the catalytic residues* in
the active site* so catalytic activity of enzyme may be lost;
8. The changes to the specific* charge of the contact/binding residues* in the active
site* may affect the temporary binding between the enzyme and substrate no
enzyme-substrate complex* formed thus may affect the catalytic process itself;

c) Explain the effect of non-competitive inhibitors on enzyme activity. [4]

No inhibitor
With non-competitive inhibitor
Rate of reaction

[S]

1. Draw and annotate a graph of uninhibited and non-competitively inhibited reaction.

2. Non-competitive inhibitors binds to allosteric site* which is another site on enzyme that
is not the active site.
3. Alters the shape/conformation of the specific* enzyme active site* so that substrate
cannot bind in correct orientation so the rate of reaction is reduced (no catalysis);
4. Rate of reaction decreases with increasing inhibitor concentration and this cannot be
neutralized by increasing the substrate concentration;
5. Some inhibitors bind reversibly to the enzyme via weak bonds such as hydrogen bonds
whilst other bind irreversibly via strong covalent bonds.

N09P2Q9 (OUT OF SYLLABUS)

Raffles Institution Nov 2009 (H2 Biology) Paper 3 (for 9744 syllabus)

Nov 2009 H2 Bio Paper 3

N09P3Q1

(a) (OUT OF SYLLABUS)

b) Explain how the ability of Taq DNA polymerase to function at high temperatures is an
advantage in the polymerase chain reaction (PCR). [5]

1. In the first step of PCR, the double stranded DNA template has to be heated to high
temperatures of about 95°C* to break the hydrogen bonds and denature the DNA;
2. Taq polymerase is not denatured at this high temperature;
3. as it is thermostable and so the first step can proceed;
4. Taq polymerase, thus can be reused after every cycle of the PCR is completed;
5. This allows the PCR to be automated;
6. The ability of Taq polymerase to withstand high temperatures also allows primer
extension to occur rapidly, and so DNA amplification can occur within a relatively
shorter time.
[Total: 15]

N09P3Q2 .

(a) A procedure for producing mammalian embryonic stem (ES) cells is shown in Fig. 2.1.

(i) Describe the characteristics of a stem cell. [2] 3L

1. A stem cell is an undifferentiated/unspecialized* cell capable of undergoing
proliferation* and self-renewal*;
2. and retains potential to differentiate* to produce specialized cells upon receiving
appropriate molecular signals*;

(ii) Explain why each cell of an 8-cell mammalian embryo, as shown in Fig.2.1, must
have the characteristics of a stem cell. [3] 5L
1. Cells up to the 8-cell embryo remains totipotent*;
2. Where each cell has ability to divide* and differentiate* for growth and form an
entire organism/foetus on its own;
3. forming all* cell types in an organism including extraembryonic tissue*;
4. This is a result of differential switching on of genes which occurs when appropriate
molecular signals are received;

(b)
(i) With reference to Table 2.1, compare the effects of the different treatments given to
the dogs in groups A and B. [2] 3L
1. Levels of expression of normal allele for G is higher in group A compared to group B;
2. There is improvement in health of 2 dogs in group A but none in group B exhibited
improvement to health, indicating that high levels of expression of allele G was
required for improvement to health;

(ii) Suggest why the expression of the normal allele of ‘G’ is different in the stem cells
given to the dogs in groups A and B. [3] 5L
1. Stem cells from an unaffected donor in group A is likely to have 2 copies of normal
functional allele G in its genome, while transduced stem cells in group B are likely to
have at most 1 copy of normal functional allele G integrated into its genome resulting
in a lower level of expression;
Raffles Institution Nov 2009 (H2 Biology) Paper 3 (for 9744 syllabus)

2. Normal functional allele G in group B may be linked to, and is regulated by a weaker
promoter* as compared to that in group A;
3. Normal functional allele in transduced stem cells in B may not be stably integrated*
into genome and hence is gradually lost with each subsequent cell division while
stable integration is not an issue with stem cells from group A;
4. Due to low success rate of gene therapy, there were much fewer successfully
transduced stem cells that were injected into group B as compared to group A;
5. Normal functional allele G for stem cells in group A was under influence of an
enhancer* element. This ensures high level of expression of normal functional allele
G as compared to group B where normal functional allele was integrated at a position
not under influence of an enhancer element;
6. Normal functional allele for stem cells in group B was integrated at a position that put
it under the influence of a silencer* element. This result in a lower level of
expression of normal functional allele G as compared to group A;
Any other valid point of comparison

(iii) Suggest one advantage of using a viral vector to add the normal allele of ‘G’ to stem
cells, rather than a non-viral gene delivery system. [1] 3L
1. Specific targeting of virus to specific cell types using specific receptors;
2. Greater efficiency of transduction of cells as viruses possess a mechanism for
attaching and entering cells;
3. Greater chances of integration* of allele into stem cell’s genome so that long-term
expression is possible. E.g retroviruses possess enzyme integrase that facilitates this
process;
Accept any one of the above 3 pts.

(c) Outline four factors that prevent gene therapy from becoming an effective treatment for
genetic disease. [4] 2L for each point.
1. Therapy is typically short-lived because of difficulty in integrating normal functional allele
into host cell genome;
2. Natural death of treated cells resulting in loss of normal functional allele. This means that
new cells need to be constantly treated through repeated treatment;
3. Many diseases are a result of multiple genes and/or multiple factors (e.g. Alzheimer’s
disease, diabetes etc) and as such will be difficult to treat with gene therapy which
currently treats only single gene disorders;
4. Some diseases such as thalassemia require a precise level of expression of normal
functional allele in order for the therapy to be effective. Regulation of levels of gene
expression is currently difficult; (Note: Not all diseases are sensitive to levels of
expression)
5. It is limited by gene delivery systems available. Host specificity of viruses can be
exploited to target specific cells but that also means that an effective viral vector may not
work if non-host cells are to be targeted;
6. If virus vector triggers immune response* the first time, immune system will mount a
stronger and faster response the second time rendering the therapy ineffective
subsequently;
7. Insertional mutagenesis* caused by retroviruses may cause inactivation of tumour
suppressor genes and/or activation of proto-oncogenes, leading potentially to cancer;
Any 4 of the above points. Note: some points from the notes not included because they are
considered minor. [Total: 15]
Raffles Institution Nov 2009 (H2 Biology) Paper 3 (for 9744 syllabus)

N09P3Q3
1. N09P3Q3
a) Suggest why it is important that Xa1 in the rice plants is expressed only in the
presence of X. oryzae. [2]
Xa1 is expressed in cells in which X. oryzae is present
1. to prevent the infection from spreading to other healthy uninfected cells of the leaf;
2. to ensure that only infected leaf cells die and not other healthy cells
3. If Xa 1 was expressed all the time then all the leaf cells will turn brown and the entire
plant will die;

b) Suggest how this duplication may account for the lack of expression of Xa2. [2]
1. The duplication would prevent general transcription factors and RNA polymerase
from recognizing and binding to the region of the allele that controls
transcription/promoter;
2. Thus preventing formation of transcritption inititation complex and hence preventing
transcription and hence preventing the production of the protein;

c) Explain how gel electrophoresis can be used to distinguish between Xa1 and Xa2. [4]
1. Since Xa2 has a 25 bp duplication, it is longer than Xa1 by 25 bp;
2. The agarose gel matrix made of a meshwork of polymer fibres which impedes
movement of longer Xa2 fragments more than shorter fragments;
3. The longer Xa 2 fragments thus migrate more slowly compared to shorter fragments,
leading to a banding pattern observed on the gel;
4. Hence the distance travelled by the Xa2 fragment from the well will be less than then
Xa 1 fragment and the banding patterns seen will be different for Xa2 and Xa1;

d) With reference to Fig. 3.1, describe and explain

i) the results of infecting the three groups of rice plants with X. oryzae [3]
1. Infection of susceptible plants, Q increased during the 4 days, reaching a value of 20
mean number of colonies of X. oryzae per leaf, since the plants do not have the Xa1
allele;

2. Infection of plants P and R increased for 3 days and reached a maximum of 10 mean
number of colonies per leaf for P and 8 mean number of colonies per leaf for R and then
decreased to 8 mean number of colonies per leaf for P and 5 mean number of colonies
per leaf for R by the 4th day;

3. Both P and R plants have resistance against X. oryzae and hence the fall in the mean
number of colonies was seen in both P and R after 3 days.

4. There was a lag time (3 days) before the numbers of bacterial colonies fell in plants P
and R because time was needed for the transcription and translation of sufficient Xa1
gene products to elicit the necessary responses (e.g. necrosis of infected leaf tissue)
and the resultant decrease in the number of bacterial colonies per leaf;

ii) the benefits of this genetic engineering of rice [2]

1. Allows for better rice yields despite presence of X. oryzae;
2. Chemicals which are harmful to the health and may leech into the ecosystem need
not be sprayed to manage the disease;
3. Allows previously blight-susceptible rice varieties to be grown in areas with a
prevalence of X. oryzae thus increasing the area and range for cultivation of those
rice varieties;
Raffles Institution Nov 2009 (H2 Biology) Paper 3 (for 9744 syllabus)

iii) the ethical implications of this genetic engineering of rice. [2]

1. Original rice plants did not contain the Xa1 allele could be cheaper than the new GM
rice thus making it unaffordable for farmers to grow and consumers to buy;
2. The long term effects of eating GM rice on human health is not known;

[Total: 15]

N09P3Q4 (OUT OF SYLLABUS)

Raffles Institution Nov 2010 (H2 Biology) Paper 2 (for 9744 syllabus)

Nov 2010 H2 Bio Paper 2

N10P2Q01
Fig. 1.1 is an electron micrograph of part of an animal cell.
(a) Identify the organelles labeled A and B.
In each case, state two visible features that enabled your identification. [6]
A: Mitochondria;
double membrane;
highly infolded inner membrane – cristae;
B: Rough* endoplasmic reticulum;
Ribosomes* studded on surface of the membrane;
flattened cisternae* comprising single membrane;

(b) (i) State the main function of each organelle. [2]

A: ATP synthesis*;
B: Protein synthesis* by ribosomes studded on membrane;
form vesicle to transport protein;

(ii) Explain how the functions of the two organelles, A and B, are linked. [2]
(A mito; B RER)
ATP produced in Mitochondria provided energy for protein synthesis;
formation of vesicles;

(c) Suggest two advantages to eukaryotic cells of membrane-bound organelles. [2]

1. Compartmentalization* maintain optimal conditions for enzyme reactions for each
organelle
2. Increase surface area for reactions (cristae for oxidative phosphorylation / attachement
ribosomes);

N10P2Q02
Fig.2.1 shows a diagram of DNA replication.
(a) Explain why the newly synthesized strand is formed continuously from the leading strand
template while Okazaki fragments are formed using the lagging strand template. [2]
1. DNA polymerase synthesizes DNA in 5’ 3’ direction* only as it will add nucleotides to
the free 3’OH group;
2. Since the two parental strands DNA are antiparallel, the daughter strand is synthesized
continuously using the 3’ to 5’ leading strand template and in short Okazaki fragments
using the 5’ to 3’ lagging strand template.

(b) Explain why this replication is said to be semi-conservative. [2]

1. Each parental strand in a DNA molecule serves as a template* for the synthesis of a
complementary* DNA strand;
2. Daughter DNA molecule comprise 1 parental (original) strand and 1 newly synthesised
strand;

(c) List three types of molecules that are required for DNA replication, apart from the molecules
shown in Fig.2.1. [3]
Helicase, Topoisomerase, Single-stranded binding proteins;
R DNA polymerase; DNA primer and RNA primase
A ATP, primase, RNA primer, free dNTPs, DNA ligase
Raffles Institution Nov 2010 (H2 Biology) Paper 2 (for 9744 syllabus)

(d) Describe how gene mutations may occur during replication of DNA. [2]
Substitution* mutation one nucleotide base is replaced* by another;
Insertion* of nucleotide bases into the original sequence
Deletion* of nucleotide bases from the original sequence

(e) Describe the effects of a mutation of one named gene. [4]

1. Sickle cell anaemia*
2. result of a single base substitution* in the gene which codes for the β-globin chain*;
CTC CAC (i.e. T is substituted by A) on the template strand,
3. 6th triplet codon* is changed from GAG to GUG;
4. codes for the amino acid valine* which is non-polar and hydrophobic instead of amino
acid glutamate* which is charged and hydrophilic;
5. at low oxygen concentration, polymerisation of Hb S sickle cell RBC
6. Sickle red blood cells are more fragile break more easily shortage of red blood
cells & poor oxygen transport;

N10P2Q03
Fig. 3.1 is an electron micrograph of a T4 bacteriophage.
(a) Identify the structures labeled A,B and C. [3]
A: Icosahedral Capsid head
B: Contractile tail sheath
C: Base plate

(b) Describe how the viral nucleic acid, contained in A, enters a bacterial cell. [3]
1. When T4 bacteriophage attaches to receptors on the bacterial host cell wall,
2. It will release lysozyme*, an enzyme which digests the bacterial cell wall resulting in a
change in the base plate conformation.
3. this in turn causes the tail sheath to contract and thrust the hollow tube through the
bacterial cell wall. (R bacterial cell membrane)
4. Viral genome will then be injected into the bacterial host cell via the hollow tube.
Raffles Institution Nov 2010 (H2 Biology) Paper 2 (for 9744 syllabus)

(c) The reproductive cycle of the T4 bacteriophage is a lytic cycle. Other bacteriophages have a
lysogenic cycle.
Explain how the two reproductive cycles of bacteriophages differ in bacterial cells. [3]
Stage T4 bacteriophage Lysogenic bacteriophage
Repli- 1. Enzymes coded for by the phage 1. Phage does not terminate the
cation genome terminates the bacterium’s macromolecular
bacterium’s macromolecular synthesis.
(DNA, RNA, protein) synthesis.

2. Phage incorporates its genome

2. Phage does not incorporate into the host cell DNA. Virus is
genome into host cell DNA. now called a prophage*.

3. No repressor protein is encoded 3. Prophage* gene expresses two

or synthesised. repressor proteins which block
expression of other phage genes
involved in phage replication.
4. Phage synthesize enzymes and
phage components immediately 4. Phage synthesize enzymes and
after infection./ There is no latent phage components after the
stage as the virus replicates once induction event in which
inside the host cell prophage is excised from host
genome and phage genes are
activated /Phage undergoes a
latent stage where it does not
undergo replication

(d) Fig. 3.2 shows the structure of tobacco mosaic virus (TMV).
List two ways in which the structure of tobacco mosaic virus:
(i) is similar to the T4 bacteriophage, [2]
1. The viral genome* is enclosed by a protein capsid*.
2. Capsid is made up of several individual protein subunits (capsomeres)
(ii) differs from the T4 bacteriophage. [2]
1. The viral genome of TMV is RNA* whilst that of T4 bacteriophage is DNA*
2. The TMV has a helical shape whilst that of T4 bacteriophage is a complex shape
consisting of an icosahedral head, collar, tail and tail fibres.

N10P2Q04
Fig. 4.1 shows a short length of chromatin.
(a) Name the structures labelled A and B. [2]
A: Nucleosomes
B: DNA
Raffles Institution Nov 2010 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Explain why the genome of eukaryotes is packaged in the way shown in Fig. 4.1. [3]
1. To make long molecule of DNA more compact to fit in nucleus*;
2. To prevent entanglement, hence prevent DNA breakage or damage;
3. Regulation of gene expression / transcription*; DNA wound around histones
prevents/hinders transcription factors and RNA polymerase access to genes that are not
meant/needed to be expressed;

(c) The histone tails of different regions of chromatin are covalently modified during the
differentiation of cells in eukaryotic organisms. These modifications are added and removed
by enzymes. The main effect of modifying histone tails is to attract specific proteins to a
stretch of chromatin.
Fig. 4.2 shows the action of one of these enzymes, histone methyl transferase.
Suggest, using Fig. 4.2 and the information above, how histone methylation prevents
transcription. [2]
1. Proteins bind methylated histones, cause chromatin* to be further compacted /
forming tighter nucleosomes;
2. Prevents binding of transcription factors* and RNA polymerase* to promoter* / to
form transcription initiation complex*, hence preventing transcription;

(ii) Suggest why histone methylation occurs over large areas of chromatin during the
differentiation of any one type of cell. [3]
1. Most of the chromatin comprises of many genes coding for proteins that are not
required in differentiated cells;
2. Hence histone methylation occurs on coding regions containing genes coding for
proteins that are not required in the cell to tightly package DNA into heterochromatin,
preventing expression of such genes;
3. A large percentage of the genome is non-coding regions and histone methylation
occurs on these regions;
4. Genome is of a large size, comprising both coding and non-coding regions;

(iii) Outline how transcription is prevented in prokaryotes during the regulation of

transcription. [3]
1. Genes coding for proteins involved in same biochemical pathway usually clustered
together on one operon*. (Expression of these genes is regulated by the same
operator and same promoter);
2. Repressor* protein binds to the operator*;
3. which prevents RNA polymerase* from binding to promoter*, thereby preventing
gene transcription;

N10P2Q05
A cross was made between two varieties of tobacco, Nicotiana, with short-tubed corollas and
long-tubed corollas.
(a) State the term used to describe the range of phenotypes in the second generation (F2) of
offspring. [1]
Continuous variation
Raffles Institution Nov 2010 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Explain why there is a range of phenotypes for this characteristic. [4]
1) Range of phenotypes are due to slight phenotypic differences that vary along a
continuum indicating continuous variation*;
2) The range of phenotypes indicate polygenic* inheritance where multiple genes are
involved;
3) There is an additive effect of each gene where each gene has a small overall effect;
4) In addition to genotypic factors, environmental factors affect the phenotype;

(c) Suggest why the range of variation is greater in the second generation (F2) of offspring than
in either the first generation of offspring (F1) or the parental phenotypes. [4]
1) During prophase I of meiosis, crossing over occurs, at chiasmata regions, between non-
sister chromatids of homologous chromosomes;
2) This results in new combinations of alleles on chromosomes;
3) During metaphase I and anaphase I of meiosis: independent assortment & random
separation of homologous chromosomes also occurs, further increasing genetic
variation;
4) This results in different combinations of maternal and paternal chromosomes in
gametes;
5) Also, if there are multiple alleles for each gene, this could lead to many more different
combinations of alleles in gametes in the F2 generation as compared to the F1 or
parental generations.
6) Random fusion of a larger variety of gametes gives rise a increased variation amongst
the F2 offspring.

N10P2Q06
Fig. 6.1 represents the main sequence of events in oxidative phosphorylation.
(a) Describe
(i) the source of the electrons at A [2]
1. Reduced NAD & FAD
2. from glycolysis, link reaction & Krebs cycle;
(ii) the fate of the electrons at B [2]
1. Combine with final electron acceptor, O2,
2. And H+ to form H2O;
(iii) the role of the high concentration of protons at C. [2]
1. Creates electrochemical proton/H+ gradient across membrane:
2. flow of H+ across the membrane down electrochemical gradient via ATP
synthase; Coupled to phosphorylation of ADP to ATP;

(b) Explain how the flow of electrons from A to B results in the high concentration of protons
at C. [3]
1. Electrons passed along electron carriers in electron transport chain in increasing order of
electronegativity;
2. Energy lost by electrons coupled to active transport of H+ against a concentration
gradient;
3. From matrix to intermembrane space;
Raffles Institution Nov 2010 (H2 Biology) Paper 2 (for 9744 syllabus)

(c) List three ways in which photophosphorylation differs from oxidative phosphorylation. [3]
Photophosphorylation Oxidative phosphorylation
Source of energy Energy for synthesis of ATP comes Energy for the synthesis of
ultimately from light. ATP comes from the
oxidation of glucose which
stores chemical energy.
Electron donors Water is the electron donor in the non- NADH and FADH2
cyclic pathway while Photosystem I is
the electron donor in the cyclic pathway.
Electron acceptors NADP+ is the final electron acceptor in Oxygen is the final electron
the non-cyclic pathway while acceptor (and it combines
Photosystem I is the final electron with H+) and is reduced to
acceptor in the cyclic pathway. water.
By product O2 H2O

N10P2Q07
Fig. 7.1 shows the distribution of four species of extinct giant flightless birds whose fossil skulls
have recently been found on five of the Hawaiian islands. The age of the islands is shown in
million years (My).
(a) Explain why evolution of different species of these birds on each island supports Darwin’s
theory of natural selection. [2]
Geographical isolation no interbreeding prevent gene flow between populations of different
islands;
Different islands different environment different selection pressures select for different
traits that are best suited to the environment;

(b) (i) Suggest why it is not possible to determine the phylogeny of the birds from the
morphological features of the fossil skulls. [2]
1. The fossil skulls were incomplete/damaged especially Chelychelynechen quassus
and Thambetochen xanion;
2. There are very few morphological characters that they all share (with just the
incomplete skull fossil) that can form a basis of comparison and so we can’t draw
any evolutionary relationships between the 4;

(ii) Explain the advantages of molecular methods in determining phylogeny. [2]

1. They are objective. Molecular character states are unambiguous as A, C, G and T
are easily recognisable and cannot be confused;
2. Data is quantitative and easily converted to numerical form for statistical analysis.
The degree of relatedness can be inferred and quantified by calculating the
nucleotide differences between species;

Context specific advantages

3. Furthermore the mtDNA does not undergo recombination thus any changes to DNA
is due solely to the accumulation of mutations over time making it the ideal candidate
for a molecular clock. We can thus estimate the time of speciation;
4. Even dead tissue may be used so long as the DNA or protein remains intact and you
do not even need an entire specimen;
(Any 2)
Raffles Institution Nov 2010 (H2 Biology) Paper 2 (for 9744 syllabus)

(c) Suggest why there are no giant flightless birds on Hawaii. [2]
1. Flightless birds will have difficulty crossing the sea to reach Hawaii from the older islands;
2. Flightless birds became extinct before the emergence of Hawaii;
3. Hawaii island emerged from the ocean some 0.43mya leaving insufficient time for flightless
birds to evolve from birds that could fly;

N10P2Q08
(a) Describe how the prokaryote, E.coli, is able to respond to varying concentrations of lactose.
[6]
1. E. coli responds to lactose through its lac operon system
2. β-galactosidase and permease are constitutively produced in the cell
3. permease (a membrane transport protein) transports lactose into the cell
4. lactose is converted by β-galactosidase to allolactose.
5. allolactose binds to repressor proteins inactivating it.
6. Inactivated repressor protein is unable to bind to operator, promoter is unblocked and
RNA polymerase can bind to promoter and transcribe structural genes
7. the structural genes are transcribed as a single polycistronic mRNA
8. lactose is also converted by β-galactosidase to glucose + galactose

(b) Explain the role of glucagon in regulation blood glucose concentration in humans. [8]
1. When blood glucose level falls below set point 90mg per 100 cm3 (stimulus),it is detected
by alpha cells of islets of Langerhans* (receptor/ detector) which also serves as
control centre
2. secrete glucagon into bloodstream;
3. Glucagon is transported via the bloodstream and binds to its specific receptor on its
main target cells i.e. liver cells (hepatocytes) (effector);

Glucagon stimulates enzymes involved in:

4. conversion of glycogen to glucose (glycogenolysis) in the liver;

5. glucose synthesis from from non-carbohydrate precursors, such as pyruvate, amino
acids and glycerol (gluconeogenesis)
6. glycogenolysis /breakdown of glycogen to glucose (ONLY in the liver);
7. break down of lipids in adipose tissues;
8. The increase in the blood glucose levels serve as negative feedback* in the form of a
diminished stimulus to the alpha cells of the islets of Langerhans of the pancreas*
and serve to decrease glucagon secretion;
9. until blood glucose level return to set point;
Raffles Institution Nov 2010 (H2 Biology) Paper 2 (for 9744 syllabus)

(c) Outline the main stages of cell signalling. [6]

3 main steps:

1. ligand-receptor interaction (reception)

2. signal transduction (inclusive of phosphorylation and signal amplification)
3. cellular responses

ligand-receptor interaction (reception)

1. a chemical signal is “detected” when signal molecule (ligand) that is structurally

complementary in conformation* binds to the ligand binding site of a specific*
receptor protein located at cell’s surface (or inside cell);
Signal transduction

2. Binding of a signal molecule (ligand) causes a change in receptor protein conformation;

3. In the case of a transmembrane protein, binding of ligand in extracellular domain of
receptor will result in a conformational change in intracellular domain* of receptor;
4. Signal transduction is initiated when signal is converted to a form that can bring about a
specific response;
Phosphorylation cascade

5. Signal cascade is often mediated by kinases and phosphatases

: Transduction of signal occurs when kinase is activated and will go on to activate a relay
protein (usually another kinase) in the next step by phosphorylating it;

6. Phosphatase will inactivate kinase in preceding step to prevent the constitutive

transduction of signal;
Signal amplification

7. this occurs via a signaling cascade;

8. At each catalytic step of cascade, number of activated products is much greater than at
preceding step;
9. Hence during transduction, signal amplification occurs too.
Cellular responses

10. Transduced signal finally triggers a specific cellular response;

o Give some examples
o e.g. catalysis by an enzyme (e.g. glycogen phosphorylase)
o physical response - rearrangement of cytoskeleton
o activation of specific genes in nucleus
11. Cell signalling ensures activities like these occur in right cells, at right time, in proper
coordination with other cells of the organism
Raffles Institution Nov 2010 (H2 Biology) Paper 2 (for 9744 syllabus)

N10P2Q09
(a) Describe the structure of an amino acid and how a peptide bond is formed with another
amino acid. [6]

1. Amino acid have 1 α-carbon atom, covalently bonded to 1 carboxyl group, 1 amino
group, 1 H atom and 1 variable R-group;
H
C O aminoH C
N group

2. R group can be a polar, non-polar, acidic or basic side group;

amino
group
OHc R-group
R a
r
b
o
x
y
l
g
r
o
u
p

1. Amino acids undergo condensation* reactions to form polypeptides with the removal of
1 water molecule with for every 1 peptide bond formed;
2. OH group from the carboxyl group and H atom from amino group contribute to the
formation of water molecule (showing OH and H in diagram coming together);
3. Box up and label peptide bond;

4. This reaction is catalysed* by an enzyme, peptidyl transferase*, at ribosomes*;

Raffles Institution Nov 2010 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Explain what is meant by primary, secondary, tertiary and quaternary structure of a named
protein. [8].
1. Named protein = Haemoglobin;
2. Primary structure refers to the unique sequence and number of amino acids in a
polypeptide linked by peptide bonds*;
3. Secondary, tertiary and quaternary structures are hence direct consequences of primary
structure;

4. Secondary structure refers to the regular coiling and folding/pleating of the polypeptide
held by hydrogen bonds* between C=O and N-H groups of the polypeptide backbone;
5. α-helix is single polypeptide wound into a coil with turns linked by hydrogen bonds*
formed between C=O* group of one amino acid residue and N-H* group of another
amino acid residue, four amino acids away;
(since this example here is haemoglobin there is no beta pleated sheets)

6. Tertiary structure refers single polypeptide chain further extensively folded and bended
into specific 3D conformation, held by bonds between R-groups within same
polypeptide;
7. Tertiary structure is maintained by hydrophobic interaction, hydrogen bonds, ionic
bonds; (in case of haemoglobin there is no disulfide bonds)

8. Quaternary structure involves more than 1 polypeptide held together by the same types
of interactions/bonds; (in case of haemoglobin there is no disulfide bonds, thus only 3
types instead of 4 types of intermolecular interactions)
9. each Hb has 4 polypeptides: 2 -globin* subunits and 2 -globin* (i.e. each polypeptide
has 1 haem group with it);
Every level of organisation of protein structure needs to be included in the answer to gain full
credit.

(c) Outline how the structure of a named globular protein is related to its specific function. [6]
1. Haemoglobin;
2. 4 polypeptide subunits: 2 -globin* subunits and 2 -globin* subunits;
3. each subunit is arranged so that most of its hydrophilic amino acid side chains are on
external surface while its hydrophobic amino acid side chains are buried in interior;
4. makes it soluble in water/aqueous environment, can be transported and carry O2 from
lungs to tissues vice versa;
5. each subunit is made of globin polypeptide and a prosthetic (non-protein) component
called haem group*;
6. each haem group consists of a porphyrin ring* and an iron ion (Fe2+)*;
7. Fe2+ of haem group binds temporarily to O2, so 1 Hb molecule can carry up to 4 O2, at a
time forming oxyhaemoglobin; (ref. transport oxygen in blood)
8. 4 subunits held together by intermolecular interactions formed between R groups
(hydrogen bonds, ionic bonds and hydrophobic interactions), allows movement that
influences affinity for oxygen allowing for cooperative binding* of oxygen;
9. As a result binding of one oxygen molecule to one haemoglobin subunit induces a
conformational change in remaining 3 subunits so that their affinity for oxygen increases;
(points 1-3, 4-6, 7-8 are 3 sets of structure-to-function relationships)
Raffles Institution Nov 2010 (H2 Biology) Paper 3 (for 9744 syllabus)

Nov 2010 H2 Bio Paper 3

N10P3Q01 (OUT OF SYLLABUS)

N10P3Q02
Rag2 mutant mice are homozygous for a mutation which causes severe combined
immunodeficiency (SCID).
The steps of the experiment are shown in Fig. 2.1.

(a) With reference to Fig. 2.1,

(i) Explain why a nucleus taken from a cell from the Rag2 mouse was used to produce the
ES cells in this therapy (step 2), [3]
1. The nucleus taken from the cell of the tail tip is diploid* while the nucleus removed from the
oocyte is haploid*;
2. By replacing the nucleus in the oocyte with the nucleus of the cell of the tail tip, this would
ensure that the embryonic stem cells produced from the oocyte would be genetically
identical* to the cells of the Rag2 mouse;
3. Therefore, these embryonic stem cells would not be rejected / evoke an immune response
when they are injected into the Rag2 mouse;

(ii) Describe the features of ES cells (step 4) and their roles in a normally developing
embryo, [3]
1. ES cells are pluripotent*;
2. Where each cell has the ability to differentiate* and divide and form an entire
organism/fetus on its own;
3. forming all* the cell types in an organism except the extraembryonic tissue*;
4. This is as a result of differential switching on of genes upon receiving molecular signals;

(iii) Explain why it is sufficient to insert a single normal allele of the Rag2 gene into each ES
cell (step 5). [2]
1. The normal allele was dominant*;
2. Expression of this allele produces a functional protein*, that mask* the effects of the
recessive mutant Rag2 allele;

(b) Targeted gene replacement of one of the mutant Rag2 alleles (step 5) is not very efficient,
but it is considered to be safer than using a viral vector to introduce the allele.
Describe the problems that may arise when using a viral vector to introduce an allele into a
cell.[3]

1. Capsid of virus poses a limit on size of normal functional allele that can be delivered to
target cells;
2. Expression of normal functional allele may be transient as not all viruses have ability to
integrate normal functional allele into target cell genome, and the normal functional allele is
therefore degraded by nucleases;
3. Random insertion of normal functional allele into genome may lead to insertional
mutagenesis*, whereby tumour suppressor genes are inactivated and/or proto-oncogenes
are upregulated and converted to oncogenes, leading potentially to cancer;
4. Virus may regain virulence once it is in host and may cause disease;
OR

1
Raffles Institution Nov 2010 (H2 Biology) Paper 3 (for 9744 syllabus)

evoke immune response which makes subsequent repeated therapy not possible as
virus would be destroyed by host immune system before it gets a chance to deliver
normal functional allele into target cells;

(c) If the procedure shown in Fig. 2.1 were to be used as a gene therapy for SCID in humans,
it might be necessary to use cow or rabbit oocytes to produce ES cells. The ethics of using
such 'cybrid' cells, in which the nuclear DNA is human but the mitochondrial DNA is from
the cow or rabbit from which the oocyte was taken, is subject to much debate.
State one argument in favour of the use of cow or rabbit oocytes for therapeutic cloning of
human cells and one argument against such use.

Argument in favour of the use of ‘cybrid’ cells [1] 3L

1. Cow or rabbit oocytes are more easily attainable than human oocytes / human oocytes
are in shortage or are limited in supply as compared to cow or rabbit oocytes;
2.
Argument against the use of ‘cybrid’ cells [1] 3L
1. Mitochondrial DNA of cows or rabbits is different from that of humans, long term effects
ofthe mixture of DNA from two different species are not predictable;
2. Not all of proteins required for proper functioning of mitochondria are encoded for by
mitochondrial DNA – some are encoded for by genes on nuclear DNA. Due to
differences between species, nuclear DNA from humans may not be able to support full
functioning of mitochondria that are of cow/rabbit origin;
Any valid point

N10P3Q03
Short tandem repeats (STRs) are regions of non-coding human DNA in which a sequence of
two to five bases is repeated, for example: GATCGATCGATCGATCGATC. The number of
these repeats, and hence the length of the STR, differs in different individuals. These different
lengths can be determined by gel electrophoresis.
(a) Explain briefly how gel electrophoresis can be used to show that a particular STR has
different lengths in different people. [4]

1. Cut DNA from different individuals with same restriction enzyme* (Since different
individuals have different number of repeats, hence different length of STR hence these
STRs of different lengths can be separated by gel electrophoresis)
2. Negatively-charged DNA* migrates towards the positive electrode/anode when subjected
to an electric field / current;
3. STR of different length can be separated because shorter fragments with fewer repeats
move faster / longer fragments with more repeats migrate slower compared to shorter
fragments
4. Meshwork impedes movement of longer fragments more than shorter fragments;
5. Carry out Southern blotting using radioactive probe* for STR repeat sequence followed by
visualising the banding pattern using autoradiography / x-ray film

2
Raffles Institution Nov 2010 (H2 Biology) Paper 3 (for 9744 syllabus)

(b) Forensic scientists create a DNA profile of an individual by extracting DNA from a sample
of cheek cells or hair roots and making multiple copies of the STRs from a number of
specific regions of the genome. The polymerase chain reaction (PCR) is used to make
these copies, as shown in Fig. 3.1.

With reference to Fig. 3.1, describe each of the three steps of one cycle of the PCR.
step 1 [2]

1. Denaturation step - Denaturation of ds DNA into ss DNA by heating to 95 C;

2. Weak H-bonds between bases of each strand broken due to increased molecular vibrations;
[2]
step 2 [2]
1. Primer Annealing Step - Temperature lowered to 64 C to allow for DNA primer to anneal;
2. Primer binds specifically to the 3’ ends of the ss target DNA by complementary base pairing;
[2]
step 3 [2]
1. Extension step - Taq polymerase performs synthesis of complementary DNA strand at 72 C;
2. Chain extension occurs from 3’ end of DNA primer which provides free 3’ OH required by
polymerase; [2]

c) The temperature at which the double strands of a STR separate into single strands can
be seen by using a molecular marker molecule. The marker fluoresces brightly when
attached to a double strand of DNA, but loses its fluorescence when the strands
separate into single strands. The temperatures at which the DNA strands separate for a
STR with 7 to 12 repeats are shown in Fig. 3.2.

With reference to Fig. 3.2,

(i) Describe and explain the trend shown by the graph, [4]
1. As number of repeats in STR increases from 7- 12， temperature needed to
separate the double strands increases from 53OC (below 55 OC ) 63.5OC
2. the temperature was increasing more slowly as the number of repeats increased
3. With an increasing number of STRs there would be an increase in the length of
polynucleotides
4. means larger number of H-bonds between bases of the two strands to be broken
5. Hence, more heat energy needed to break more bonds and separate strands

(ii) Suggest one difficulty that is likely to be encountered when using this technique to find
the length of STRs with larger numbers of repeats. [1]
(The temperature was increasing more slowly as the number of repeats increased). STR with
larger number of repeats beyond 12 repeats may separate at the same temperature and hence
unable to determine the number of repeats in STR

N10P3Q05 (OUT OF SYLLABUS)

3
Raffles Institution Nov 2011 (H2 Biology) Paper 2 (for 9744 syllabus)

Nov 2011 H2 Bio Paper 2

N11P2Q1
(a) Describe the shape of the curve when no inhibitor is present. [2] (4 lines)
1. From 0 to about 10 arbitrary units of substrate concentration, there was a steep/rapid initial
increase in the rate of reaction from 0 units to about 6 arbitrary units;
2. After 10 arbitrary units of substrate concentration, the rate of reaction still increases but the
rate of its increase is now decreasing;
3. The curve started to level off from 20 to 100 arbitrary units of substrate concentration. The
plateau value for the rate of reaction was 10 arbitrary units;

(b) (i) On Fig. 1.1 draw the curve you would expect in the presence of a competitive inhibitor. [2]

competitive inhibitor

1. Curve drawn between the non-competitive inhibitor and no inhibitor curves;

2. Curve must meet the no inhibitor curve at higher substrate concentrations i.e. at 100 units
substrate concentration and 10 units rate of reaction; (see diagram above)

(ii) Explain with reasons the shape of the curve you have drawn. [3] (6 lines)
1. Competitive inhibitor has a similar charge and shape/ conformation* to the substrate;
2. allowing it to bind to the enzyme active site* and block substrate binding;
3. The binding of the inhibitor to the active site is not permanent/reversible;
4. Effect of the inhibitor at high substrate concentrations is negligible since the higher
proportion of substrate molecules compared to inhibitor molecules can effectively out-
compete the inhibitor molecules for the active sites;
5. Hence at very high substrate concentrations, the rate of reaction is the same as that when
no inhibitor was present;
(c) (OUT OF SYLLABUS)

N11P2Q2
(a) Identify A – D. [4]
A: large ribosomal subunit of ribosome*
B: amino acids of growing polypeptide*
C: tRNA* (accept:transfer RNA)
D: ribonucleotides of mRNA* (accept: messenger RNA)
Raffles Institution Nov 2011 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Explain how the correct amino acid is joined to a tRNA. [4] (8 lines)
1. The enzyme is aminoacyl-tRNA synthetase*;
2. It has a specific active site* that has complementary shape/conformation and charge* to
3. a tRNA* with a specific anticodon* and;
4. a specific amino acid* to be attached to corresponding tRNA

(c) State the role of rRNA in protein synthesis. [3] (6 lines)

1. rRNA associates with a set of ribosomal proteins* to form large and small subunits of
ribosomes*;
2. small ribosomal subunit can bind to mRNA* via complementary base pairing* whereby
adenine base pairs with uracil and guanine base pairs with cytosine.
3. large ribosomal subunit enables binding of aminoacyl-tRNAs* to P site (peptidyl-tRNA
binding site) and A site (amino-acyl tRNA binding site)
4. rRNA molecule forms peptidyl transferase* on large ribosomal subunit which catalyses
formation of peptide bond* in the polypeptide

N11P2Q3
(a) Suggest the functions of haemagglutinin and neuraminidase in an enveloped virus, such
as influenza. [2] (2 lines each)
1. Haemagglutinin enables the virus to bind/attach to the sialic acid receptor at the plasma
membrane of the host cell in order to gain entry into it;
2. Neuraminidase facilitates the release of the newly formed virus from the host cell during the
budding process.

(b) Other than haemagglutinin and neuraminidase, state two other components of the envelope
of a virus, such as influenza. [2]
1. phospholipid bilayer
2. glycoproteins
3. cholesterol

(c) Explain how an enveloped virus, such as influenza, enters a cell. [4] (8 lines)
1. Haemagglutinin* binds to sialic acid receptor* on host cell membrane
2. Virus then enters host cell by endocytosis*: where host cell membrane invaginates* and
pinches off, placing virus in an endocytotic vesicle*.
3. Vesicle then fuses* with a lysosome*, and the resulting drop in pH stimulates fusion of viral
envelope with vesicle membrane
4. Releasing nucleocapsid* into cytosol
(d) (i) With reference to Fig. 3.1, suggest how the new combination of RNA segments in
influenza virus P could have arisen. [2] (4 lines)
1. 3 strains of viruses - classical swine, North American Avian and human H3N2 infect the
same cell of a pig.
2. RNA segments H1,NP, M and NS from classical swine, PB2 and PA from North American
Avian and PB1 and N2 from human are assorted together in a virion* that forms a new
influenza virus P, giving rise to antigenic shift*.
Raffles Institution Nov 2011 (H2 Biology) Paper 2 (for 9744 syllabus)

(ii) Influenza virus Q and human H1N1 have RNA segments with the same origin but differ
in their ability to infect humans. What changes may have occurred to give human H1N1
greater ability to infect humans? [2] (4 lines)
1. The mutation* of haemagglutinin (H1) occurs such that it has a more precise binding
site, for the sialic acid receptor of the human respiratory epithelium cell to enter the host
cell.
2. The mutation* of neuraminidase (N1) such that it has a more precise active site which
facilitates the release of virions/newly assembled viruses from the host cells by budding
to re-infect other cells.

N11P2Q4
(a) Explain how this mutation may change the structure and function of ras protein. [4] (8 lines)
1. Single base substitution GGC GTC *changes 1 amino acid gly valine*
2. Different R groups affecting the type and location of bonds formed (eg: hydrogen bonds,
ionic bonds, disulphide bonds etc)
3. hence altering secondary and tertiary protein structure / 3D conformation* of ras protein*.
4. Binding site of mutated ras protein no longer complementary in shape and cannot bind to
target DNA sequences/ other proteins to influence gene expression, hence cannot act as
transcription factors;

(b) Distinguish between a gain of function mutation and a loss of function mutation. [4] (8 lines)
Compare any 2 points for gain in function, and 2 corresponding points for loss in function
Gain-of-function mutation Loss-of-function mutation
1. results in a gene product that is 2. results in a non-functional gene product/
hyperactive/ produced excessively; gene product not produced;
3. only a single mutation* is needed to have 4. need to mutate both alleles* to have
effect; effect;
5. mutation is usually dominant* 6. mutation is usually recessive*

(c) State two factors that can cause mutations. [2]

1. such as ultraviolet light / radioactivity / ionizing radiations, (mention of radiation alone not
sufficient);
2. Carcinogens such as tar in cigarette smoke, asbestos, benzene, formaldehyde, ethidium
bromide etc. (give named examples);
3. Viruses like avian sarcoma virus, HPV;

(d) Using the details provided in Fig. 4.1 and Fig. 4.2 describe how this mutation of a proto-
oncogene to oncogene differs from the mutation that involves the ras protein. [2] (4 lines)
1. Fig. 4.2 shows in Burkitt’s lymphoma, chromosomal mutation where a long DNA stretch
/portion of chromosome (c-myc) from chromosome 8 is transferred to chromosome 14,
whereas in mutation of ras protein, Fig. 4.1, shows a gene mutation where one nucleotide
base is substituted;
2. In Burkitt’s lymphoma, Fig. 4.2’s regulation of gene expression is altered, producing the
same gene product whereas, in mutation of ras protein, Fig. 4.1 regulation of gene
expression is not affected, rather a different gene product is produced.
Raffles Institution Nov 2011 (H2 Biology) Paper 2 (for 9744 syllabus)

(e) Explain why each of the homologous chromosomes appear as double structures in Fig. 4.3.
[2] (4 lines)
1. DNA replication* takes place during S phase of interphase (before cell division) producing 2
molecules of DNA.
2. chromosomes become visible as they condense* and shorten prior to cell division during
prophase so that each chromosome consists of two sister chromatids* joined at the
centromere.

(f) Describe the different effects of these two oncogenes in causing tumours in mice. [3] (6 lines)
1. Mice with a single c-myc mutation don’t develop tumours as easily as with a single H-ras
mutation. E.g: at day 200 60% c-myc mutants vs 40% H-ras mutants.
2. Mice with a single c-myc mutation develop tumours later than those with a single H-ras
mutation. E.g: c-myc mutants begin at day 100 vs H-ras mutants begin at day 50.
3. Mice with double mutations have the lowest survival rate, none survived at day 200.

N11P2Q5
(a) Explain the meanings of the terms
(i) locus, [2] (4 lines)
The position of a gene or allele on the chromosome
(ii) allele. [2] (4 lines)
1. Allele is the alternative form of a gene
2. Alleles of the same gene are found on the same loci of the homologous chromosomes
3. They may be recessive, dominant or codominant
Raffles Institution Nov 2011 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Draw a genetic diagram showing the cross between Beatrice and Henry. For each of their
children, state their possible genotype(s) and phenotype. [5]

Let XH represent the female chromosome carrying the normal allele for blood clotting (dominant)
Let Xh represent the female chromosome carrying the allele for haemophilia (recessive)
Let Y represent the male chromosome

Normal female (carrier) Normal male

Parental phenotypes (Beatrice) X (Henry)
Parental genotypes XH Xh XH Y
Meiosis
Gametes

Fertilization

Gametes

H
X Y

X
h XH Xh Xh Y
Gametes
Female carrier Haemophilic male

X
H XH XH XH Y
Normal female Normal male

Offspring genotype XH XH XH Y XH Xh Xh Y
1 Normal : 1 Normal : 1 Female : 1 Haemophilic
Offspring phenotypic ratio female male carrier male

Alexander: XH Y normal male

Eugenie: XH XH normal female or XH Xh female carrier
Leopold Arther: Xh Y haemophilic male
Maurice: Xh Y haemophilic male

[2] for parental genotypes and gametes and for punnett square
[1] for each correct genotype and phenotype of Eugenie, Alexander, Leonard & Maurice

(c) None of Queen Victoria’s ancestors suffered from haemophilia. Suggest in whom and in
what organ a mutation occurred that accounts for haemophilia in her family. [2] (4 lines)
1. Spontaneous mutation in the ovary of Queen Victoria so that the gametes produced contain
the haemophilia allele that is inherited by Leonard and Beatrice.
2. It cannot be a mutation in the testes of Prince Albert since Leonard inherits only the Y
chromosome from the father that does not carry the haemophilia allele.
Raffles Institution Nov 2011 (H2 Biology) Paper 2 (for 9744 syllabus)

N11P2Q6
(a) Explain why ions such as Na+ and K+ cannot freely pass through the phospholipid bilayer of
membranes. [2] (4 lines)
1. the hydrophobic region / core* of the phospholipid bilayer
2. prevents the charged* Na+ and K+ from passing across the phospholipid bilayer.
*NB : reject reference to Na+ and K+ as polar molecules

(b) Describe the two ways in which ions such as Na+ and K+ can pass through the membrane
shown above. [2] (4 lines)
1. These charged ions move through specific ion channels (e.g. leak, voltage-gated and
ligand-gated ion channels) embedded in the membrane of the nerve cells by facilitated
diffusion* down an electrochemical gradient*;
2. And through ion pump/ carrier protein e.g Na+-K+ / Ca2+ pump where ions are actively
transport* against their concentration gradient* with the expenditure of ATP*;

(c)-(e) (OUT OF SYLLABUS)

N11P2Q7
(a) Explain the relationship between classification and phylogeny. [3] [6 lines].
1. Classification is the organisation of species according to particular characteristics and may
not take into consideration the evolutionary relationship between the species.
2. Phylogeny is the organisation of species according to particular characteristics which takes
into consideration the evolutionary relationship between the species.
3. The relationship between classification and phylogeny is phylogenetic classification where
species are categorised into the hierarchical classification based on the evolutionary
relationships derived from phylogeny

(b) Explain why molecular evidence is better than morphological evidence in determining
phylogeny. [4] [9 lines]
[Note: Any four of the below points (as far as possible try to refer to the phylogenetic tree and
the use of rRNA above. Points 1 - 4 are the better points to highlight).]
1. All known life is based on nucleic acids* and would have rRNA and thus studies involving
any types of taxa can use rRNA sequence data;
2. The rRNA sequences can be used to compare species that are morphologically difficult to
compare to establish evolutionary relationships e.g., animals and bacteria;
3. rRNA sequences are objective and quantitative*. Molecular character states are
unambiguous* (e.g. A, C, G and U) whereas some morphological characters such as
colour, texture, can be ambiguous; (note: reject A,G, C T as the molecule used is RNA)
4. Using the molecular clock* applied to the ribosomal RNA, it can be determined when the
different kingdoms of life diverged from each other.
5. Molecular characteristics can clearly show if organisms are wrongly classified due to
morphological characters that are similar between organisms due to convergent
evolution*.
Other valid points
6. Molecular data (nucleotide or amino acid sequences) are easily converted to numerical
form* and hence are amenable to mathematical and statistical analysis.
7. Offers a large and essentially limitless set of characters* to be studied. Each nucleotide
position can be considered a character and the ribosomal RNA can provide about 5
thousand bases on average;
Raffles Institution Nov 2011 (H2 Biology) Paper 2 (for 9744 syllabus)

N11P2Q8
(a) Compare the Krebs cycle and the Calvin cycle. [6]
Features Calvin cycle Krebs cycle
1. Site Takes place in the stroma* Takes place in the matrix* of
of chloroplast mitochondria
2. Nature of process Anabolic process – involves Catabolic process - involves
the formation of G3P or the breakdown of acetyl coA
sugars (e.g. sugars) and and intermediates. It also
involves a reduction reaction. involves oxidation by
dehydrogenation.
3. Fate of carbon 1 molecule of CO2 is 2 molecules of CO2 is
dioxide accepted by RuBP catalyzed released by oxidative
4. per cycle by Rubisco decarboxylation
5. Role of Reduced NADP / NADPH + NAD+ & FAD as electron and
coenzymes H+ as electron and proton proton acceptors
involved donor
6. Role of ATP Expenditure of ATP - in the Synthesis of ATP by substrate
reduction of GP to G3P and level phosphorylation
in the regeneration of RuBP
7. Substance Ribulose bisphosphate (5C) Oxaloacetate (4C)
regenerated

(b) Discuss the roles of NAD and NADP. [6]

1. Both NAD and NADP are coenzymes which carry both protons and electrons.
2. NAD, in its reduced form (NADH), carries electrons from oxidation reactions in glycolysis (in
cytoplasm), the link reaction and Krebs cycle (both in mitochondrial matrix),
3. to the electron transport chain at the cristae.
4. NAD is regenerated when NADH is reoxidised at the electron transport to continue its role
as an electron carrier.
5. NADP is the final electron acceptor in the light dependent reaction in the thylakoid
membrane
6. Electrons carried in reduced NADP is brought to the stroma and used in the Calvin cycle in
the stroma of the chloroplast.
7. When glycerate phosphate (GP) is reduced to glyceraldehyde-3-phosphate (G3P), NADP is
regenerated to carry out its role as an electron carrier.

(c) Outline the main features of photo-phosphorylation. [8]

1. Light energy* is absorbed by accessory pigments molecules, such as chlorophylls a and
b* in Light Harvesting Complex (LHC) of Photosystem (PS) I and II;
2. These pigment molecules in LHC pass on this energy to the next pigment molecule by
resonance transfer of energy until a special chl a molecule in the reaction centre is reached;
3. When special chlorophyll a molecule absorbs the energy, an electron is excited*to a higher
energy level, Chl a chl a+ +e- , leaving an e- hole in PS I and II;
4. At photosystem II, the electron hole is filled by the photolysis of water* which donates a
pair of electrons and yields molecular oxygen as a by-product;
5. Excited electrons* flow down a chain of carriers of increasing electronegativity along the
electron transport chain* until it reaches;
6. NADP+*, which is the final electron acceptor of the non-cyclic photophosphorylation. NADP+
gets reduced to NADPH* by NADP reductase;
Raffles Institution Nov 2011 (H2 Biology) Paper 2 (for 9744 syllabus)

7. The energy loss along the electron transport chain is used to actively pump protons across
the thylakoid membranes into the thylakoid space;
8. This leads to the generation of a proton gradient* across the thylakoid membrane. As H+
moves down its concentration gradient via the ATP synthase* complex, it is used to
phosphorylate ADP to ATP*. This process is called chemiosmosis*;
9. Cyclic photophosphorylation where excited electrons are recycled back to PSI via another
chain of electron carriers, generates additional ATP* for the light-independent reaction of
photosynthesis.

N11P2Q9
(a) Compare the structure of prokaryotic and eukaryotic genomes. [6]
Feature Structure of Eukaryotic Genome Structure of Prokaryotic Genome
1 Size of genome 108-1011 base pairs 104-107 base pairs
2 Appearance of genetic Multiple, linear molecules Generally a single, circular molecule
material
3 Molecule making up Double helix DNA Double helix DNA
the genetic material
4 Association of genetic Yes – large amounts of it e.g. naked DNA, insignificant amounts
material with proteins histones, scaffold proteins
5 Level of DNA High: Relatively low:
packing/coiling DNA is associated with equivalent DNA is associated with small
mass of histone; amounts of protein;
Different levels of packing present some looping is present around
(nucleosome solenoid 30 nm nucleoid-associated proteins
fiber looped domains supercoiling of loops bacterial
chromosome) chromosome
6 Location of genetic Nucleus Nucleoid region – not membrane-
material in cell bound
7 Origin of replication Multiple Single
8 Introns present Yes No
9 Other non-coding Centromeres and telomeres present Centromeres and telomeres absent
sequence
10 Gene organization Monocistronic Organised as operons
11 Number of genes Many Fewer

(b) Discuss the role of telomeres and centromeres. [6]

Telomeres (max 4 marks)
1. Telomeres are non-coding* tandem repeat sequences
2. found at both ends of chromosomes; (non-coding must be mentioned somewhere in answer)

Role – main
3. Each round of DNA replication will result in the shortening of daughter molecules at the
telomeres because DNA polymerase is unable to replace the RNA primers with DNA; (idea
of end replication problem) [R: prevent end replication problem].
4. Since telomeres are non-coding, this ensures that vital genetic information/genes are not
lost/ eroded with each round of replication; wtte

Role – others
5. By forming a loop with 3’ overhang, they protect and stabilise terminal ends of chromosome,
hence preventing fusion of the ends with those of other chromosomes;
Raffles Institution Nov 2011 (H2 Biology) Paper 2 (for 9744 syllabus)

6. Either: prevent triggering pathways that lead to cell arrest or cell death, because exposed 3’
overhang will be perceived as DNA damage/DNA double strand break;
Or: prevent DNA repair machinery from recognizing the ends of chromosomes as DNA
breaks/ damage, hence preventing apoptosis;
7. Either: The 3’ overhang of the telomeres allow their own extension, by providing an
attachment point for the correct positioning of the enzyme telomerase in certain cells, e.g.
germ cells
Or: They possess a 3’ overhang which base pairs with the RNA template on telomerase, so
ensures proper alignment of telomerase and allows extension of telomeric ends in certain
cells e.g. germ cells.

Centromeres (max 2 marks)

1. Centromeres are non-coding DNA consisting of tandem repeat sequences found at one
location anywhere along the length of the chromosome;
2. They allow sister chromatids to adhere to each other;
3. They allow proteins called kinetochores*to attach,
4. and subsequently spindle fibres*, to attach so that sister chromatids/homologous
chromosomes can be separated to opposite poles;

(c) Outline the main features of DNA replication. [8]

1. Replication of DNA is semi-conservative* where the original strands of the double helix
separates* and act as templates* for the synthesis of two new strands;
2. This gives rise to two new/daughter DNA molecules, each consisting of one original and one
newly synthesised strand;
3. Replication of DNA begins at the origin of replication* where enzyme helicase* will bind
and unzip* the DNA molecule by breaking hydrogen bonds* between complementary base
pairs;
4. Separated strands of DNA interact with the single stranded DNA binding proteins* so that
it will remain single stranded and can serve as a template* for replication;
5. Topoisomerase* relieves overwinding strain ahead of replication forks by breaking,
swivelling and rejoining DNA strands;
6. Enzyme primase* catalyses the synthesis of a short RNA primer*; (Reject: RNA primase)
7. Complementary base pairing* occurs between template* strand and free incoming
deoxyribonucleoside triphosphates*;
8. Whereby adenine* forms 2 hydrogen bonds* with thymine* and guanine* forms 3
hydrogen bonds with cytosine*;
9. DNA polymerase* catalyses the formation of phosphodiester bond* linking DNA
nucleotides;
10. and the new DNA strand is synthesized in the 5’ to 3’ direction*; (@: template strand read
from 3’ 5’ direction*)
11. RNA primers* will then be removed and replaced by DNA by another DNA polymerase;
12. Due to the anti-parallel nature of the DNA molecule, one of the daughter strands known as
the leading strand* will be synthesised continuously towards replication fork;
13. the other strand known as the lagging strand* is synthesised discontinuously away from
the replication fork, giving rise to Okazaki fragments*;
14. DNA ligase* catalyses the formation of phosphodiester bond* between the Okazaki
fragments, sealing the nicks*;
Raffles Institution Nov 2011 (H2 Biology) Paper 3 (for 9744 syllabus)

Nov 2011 H2 Bio Paper 3

N11P3Q1 (OUT OF SYLLABUS)

N11P3Q2 (OUT OF SYLLABUS)

N11P3Q3 (OUT OF SYLLABUS)

N11P3Q4
Planning question [12]

1. AIM: To investigate the effect of light intensity on the rate of photosynthesis of mesophyll
cells from the sun and shade leaves of the Johor Fig.

2. Theory:
(How light intensity affects light dependent reaction? In turn, how does light dependent
reaction affect carbon fixation?)
a. As a higher intensity of light is absorbed by chloroplasts, there will be a more electrons
excited in chlorophyll. More electrons are emitted by PS I and II, and hence reducing
NADP. More NADPH available for the light independent reaction, more fixation of CO2.
main theory [1m]
b. As the rate of CO2 fixation increases and the time taken for the concentration of CO2 of
the bathing solution to fall from 1.18 x 10-5 moldm-3 (pH of 5.65) to 3.36 x 10-6 moldm-3
(pH is 5.92) is shorter Variable that you are measuring & expected observation
c. As light intensity increases, the rate of photosynthesis increases in both sun and shade
leaf cells. predicted trend
d. Protoplasts from shade leaf are better adapted in the shade compared to protoplasts from
sun leaf. Hence, the maximum rate of photosynthesis in shade leaves is lower compared
to that of the sun leaves. predicted trend
e. Light saturation is reached at a lower light intensity for shade leaf protoplast compared to
sun leaf protoplast. predicted trend

3. Procedure:
a. Pilot test
Conduct a pilot experiment to determine suitability of apparatus, suitable range of
independent variables, optimum conditions and amount of materials used.

b. well-Annotated diagram
Raffles Institution Nov 2011 (H2 Biology) Paper 3 (for 9744 syllabus)

neutral density
filter tube A slim pH probe inserted through
tube’s lid and immersed in
plastic tube with lid bathing solution

15cm pH 5.80 pH meter

ruler bead with protoplasts

bench light from shade leaf in
bathing solution

c. Numbered steps
Dependent variables: rate of photosynthesis
Independent variables: light intensity

1. Set up the experiment as shown in the diagram.

2. Constant variables: number of beads, distance from light source, and volume of
bathing solution are kept constant for accuracy of results. [at least 2 e.g.; must
state condition that is kept constant, describe how this is done & why this is
done in the procedure]
3. Place 3 beads of shade leaf protoplasts into a plastic tube. Using a syringe, add
5ml of bathing solution. Number of beads is kept constant to ensure constant
number of protoplasts/chloroplasts. Screw the lid of the plastic tube.
4. Then place the plastic tube into neutral density filter tube A (allows 6% light to be
transmitted).
5. Leave the tube in the dark for 2 minutes for equilibration to ensure that all the
protoplasts stop photosynthesizing.
6. Measuring with the ruler, place the tube 15cm from the bench light. Distance of
light source kept constant to ensure that the varying light intensity is due to the
different ability of filter tube to transmit light.
7. Using the stopwatch, start timing when the lamp is switched on. Record the
measurable quantity which is the time taken for bathing solution to increase from pH
5.65 to pH 5.92. The dependent variable is the rate of photosynthesis calculated by
1/time taken.
8. Repeat steps 3 to 7 twice using neutral density filter tube A. These serve as
replicates of neutral density filter tube A to check for anomalous results.
9. After doing 3 replicates, carry out steps 3 to 8 for neutral density filter tubes B to F.
Independent variable is light intensity. The tubes A to F allow different amount of
light transmitted, with 6%, 12%, 25%, 50%, 71% and 100% respectively.
10. After the entire experiment for shade leaf protoplasts, repeat the experiment from
steps 3 to 9 using beads of sun leaf protoplasts.
Raffles Institution Nov 2011 (H2 Biology) Paper 3 (for 9744 syllabus)

d. Control (1m)
Set up 2 controls, one with 3 beads of shade leaf protoplasts in 5ml bathing solution and
the other with 3 beads of sun leaf protoplasts. Keep both in the dark and measure the pH
of the bathing solutions at the end of the experiment.
This is to show that any pH change is due to the amount of light available for
photosynthesis and not due to any other factors.

e. Repeats
Repeat the whole experiment to check for reproducibility.

4. Data recording and processing

Table showing rate of photosynthesis at different light intensities in shade protoplasts

Neutral Percentage Time taken for end point (pH=5.92) to be Rate of

density of light reached/min photosynthesis/
filter transmitted/% Replicate Replicate Replicate Average min-1
tube 1 2 3
A 6
B 12
C 25
D 50
E 71
F 100

A table of similar format will be used to record the experiment for sun protoplasts.

Graph showing rate of photosynthesis at different light intensities in shade protoplasts

Rate of Sun protoplasts

photosynthesis
-1
/ min
Shade protoplasts

Light transmitted/%

5. Risks and Precautions (1m)

a. Ensure that hands are dry when switching on the lamp to prevent electrocution.
Raffles Institution Nov 2011 (H2 Biology) Paper 3 (for 9744 syllabus)

neutral density neutral density slim pH probe

neutral density filter tube B inserted through
filter tube A filter tube D
tube’s lid and
immersed in
plastic tube bathing solution
with lid
pH 5.80 pH meter

neutral neutral
density density bead with protoplasts
filter tube filter tube from shade leaf in
15cm bathing solution
C E
neutral density
filter tube F

ruler

bench light

Alternative set-up for the experiment.

N11P3Q5
Explain:

(a) How gel electrophoresis is able to separate fragments of DNA; [6]

1. Dense loading buffer mixed with DNA sample to help it sink to the bottom of the well;
2. Loading dyes also added to DNA sample to allow visualisation of the separation process;
3. The fragments of DNA are pipetted into the wells at the top of the gel in a position furthest
from the positive electrode/anode;
4. Negatively-charged DNA*;
5. Migrates out of well towards direction of positive electrode/anode when subjected to an
electric field / current;
6. Fragments migrate through agarose gel matrix, made up of a meshwork of
polysaccharides;
7. Meshwork impedes movement of longer fragments more than shorter fragments;
8. Longer fragments migrate slower compared to shorter fragments;

This question was also asked in Nov 2008/Q1a(ii). The examiner’s comment was that the
location of the wells in the agarose gel was not always clear. Students also were confused
between anode and cathode.
Raffles Institution Nov 2011 (H2 Biology) Paper 3 (for 9744 syllabus)

(b) what is meant by restriction fragment length polymorphism (RFLP) and how it can be
detected; [6]

1. Restriction Fragment Length Polymorphism (RFLP) refers to the unique banding pattern
among individuals when the genomic DNA are digested by restriction enzymes separated by
gel electrophoresis;
2. Due to the presence of DNA polymorphisms different individuals, there will be variations in
number and location of restriction sites and number of tandemly repeated nucleotide
sequences (e.g. micro and minisatellite loci) among individuals;
3. To detect polymorphisms, the DNA fragments are transferred to nitrocelluose membrane
and made single stranded using an alkaline solution;
4. nucleic acid hybridization is carried out by incubating the membrane with radioactive DNA
probe complementary to the region of interest;
5. radioactively labeled probe binds to complementary DNA fragments, appearing as bands in
the autoradiograph, highlighting the differences in the restriction sites and hence DNA
polymorphisms in different individuals;

(c) how RFLP analysis has helped the process of detecting a named genetic disease. [8]
1. Sickle-cell anaemia* is an autosomal recessive genetic disease which is caused by a
substitution mutation in the DNA coding for β-globin chain of haemoglobin, where T is
substituted by A;
2. Red blood cells assume an abnormal rigid, sickle shape and can lead to anemia, obstruction
of capillaries due to the sickling, organ damage, drops in haemoglobin levels and
subsequently death;
3. This mutation is located within a restriction site for the restriction enzyme MstII* in the
disease-causing allele (HbS) and as a result, the restriction site is no longer recognised by
MstII;
4. When normal and disease alleles are digested by MstII, a different mixture of fragments will
be produced from each allele. It is possible to infer the genotype of individuals using RFLP
analysis;
5. To detect the disease, DNA from the affected individual is digested with MstII and the
digested DNA separate using gel electrophoresis;
6. Southern blotting is carried out whereby double stranded DNA is made single-stranded
using NaOH and transferred to a nitrocellulose membrane;
7. for nucleic acid hybridization (Southern blot) using a radioactive-labeled probe that is
complementary to the β-globin gene on either side of the restriction site;
8. because the HbS allele has lost the restriction site through substitution, it will hence yield a
larger fragment when the fragments are visualized in the autoradiograph;
9. A single heavy/larger band will indicate the presence of 2 HbS alleles and hence a sufferer.
2 bands, one intermediate and one small will indicate the presence of 2 HbA alleles and
hence a normal individual. 3 bands, one large, one intermediate and one small will indicate
a heterozygote who is also normal;
Raffles Institution Nov 2011 (H2 Biology) Paper 3 (for 9744 syllabus)

Teachers’ comment:
“for a named genetic disease” I believe the examiners want you to name the disease! Please
give some details about the disease like what gene is affected, the mutation, and how it causes
disease. Details are hard to come by in most scripts. Few mentioned the alleles HbS and HbA
preferring to use “disease and normal alleles”. This is a process question so describe the
process. Simply stating “Southern blot hybridization”: is not a description of a process. Always
explain the various possible outcomes and the conclusion you can draw.(pt 8) This seems to be
a major weakness amongst students.

Short bar shows

region where
radioactive
labeled probe is
complementary to
Raffles Institution Nov 2012 (H2 Biology) Paper 2 (for 9744 syllabus)

Nov 2012 H2 Bio Paper 2

N12P2Q1

(a) Identify the stage of mitosis shown in Fig. 1.1. [1]

Metaphase*

(b) Describe what happens in the next stage of mitosis. [3]

1. During anaphase* ;
2. centromere*(s) divides/duplicates (R replicate/split) and sister chromatids*, now
chromosomes,* ;
3. pulled/separate to opposite poles* (Reject: ends ) by shortening kinetochore ;
microtubules* (Reject: contract) with centromeres leading resulting in characteristic
v shape;

(c) Outline the role of centrioles in mitosis.[2]

1. There are a pair of centrioles at each pole and their position determines the polarity of
the cells;
2. Centrioles form part of the microtubule-organising centre and are involved in organizing
the synthesis of spindle fibres;
3. Which lead to separation of chromosomes after centromere divide;

(d) Describe and explain what happens to the nuclear envelope during mitosis. [4]

1. During prophase*, the nuclear envelope disintegrate;

2. to allow the chromosomes to move to the metaphase plate;
3. During telophase*, the nuclear envelope reforms;
4. To enclose the chromosomes inside the nucleus;

(e) Suggest the specific cell structures targeted by the two fluorescent dyes used in the
preparation. Their binding is shown on Fig. 1.1 in regions A and B. [2]

Structure in region A- chromosomes aligned at the metaphase plate

Structure in region B- microtubules and asters

[Total: 12]
N12P2Q2

(a) Explain why chloride ions can only cross cell surface membranes via such proteins.[3]

1) Chloride ions are charged (R: polar);

2) The hydophobic core* of the phospholipid bilayer* of the cell surface membrane
would repel charged ions, preventing chloride from moving across the cell surface
membrane;
3) The protein channel, a transmembranal protein, provides chloride ions with a
hydrophilic pathway to move across the membrane;
Raffles Institution Nov 2012 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Describe how the membrane holds onto these proteins.[2]

1) The non-polar hydrocarbon chains of the phospholipid bilayer can form hydrophobic
interactions*;
2) with the R groups of the non-polar amino acids found on the exterior surface of the
channel protein (facing the hydrophobic core);

(c) Suggest the role of ATP in opening the channel. [2]

1) ATP binds to the ATP-binding domain of the CFTR causing a change in conformation;
2) Which leads to the opening of the pore, allowing chloride ions to move out;

(d) State the type of gene mutation that causes the commonest mutation to the CFTR gene. [1]

1) Deletion of 3 nucleotides coding for the amino acid phenylalanine;

(e) Explain how such structural changes in a protein may affect its functioning.[3]

1) The loss of an amino acid, phenylalanine, changes the *rimary* structure of the CFTR
protein;
2) The amino acid is no longer available to form bonds involved in the maintenance of the
secondary* and tertiary* structure of the protein;
3) This results in a change in the overall 3D conformation of the CFTR protein, particularly
in the ATP-binding domain;
4) This results in an inability to bind to ATP and hence, no opening of the pore to allowing
chloride ions to diffuse out;

(f) Explain why only one copy of the normal allele of the CFTR gene is required to prevent
cystic fibrosis in individuals with the most common mutation present in the other allele. [2]

1) With one normal allele, the normal CFTR protein will still be produced
2) which will allow the movement of chloride out of the cells;
3) As chloride move out, water will also move out by osmosis* which will dilute the mucus
outside the cell thus there will be no sticky or think mucus;
[Total: 13]

N12P2Q3
(a) State the name of the structures shown in Fig. 3.1.[4]

A – capsid head
B – contractile sheath
C – tail fibre
D – tail pin (R: base plate)
Raffles Institution Nov 2012 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Describe the role of viral DNA in the infection process of the T4 bacteriophage. [3]

1. Viral DNA codes for / is transcribed and translated to produce enzymes and structural
components of phage which are important for infection of bacterial cell;
2. Enzymes hydrolyse the DNA of host cell to nucleotides which can be used by host DNA
polymerase to replicate viral DNA;
3. Enzyme lysozyme* which is released by phage to digest bacterial cell wall to trigger
lysis of host cell for release of newly formed virions / to trigger injection of viral DNA into
host cell;
4. phage proteins which form tail fibres which can recognise and bind to receptors on
surface of bacterial cell;
5. phage proteins which form base plate* which can change shape in response to
molecules released from bacterial cell to cause contraction of contractile sheath for
injection of viral DNA.;

(c) Suggest the advantages of the lysogenic pathway to the lambda phage virus. [2]

1. Via the lysogenic pathway, the lambda phage DNA is integrated into the bacterial DNA
forming a prophage* ;
2. every time the host cell’s machinery replicates the bacterial chromosome, it also
replicates the prophage DNA along with it ;
3. This allows continuous replication of the lambda phage DNA/prophage without killing the
host bacteria.;

[Total: 9]
N12P2Q4

(a) Suggest how transcription factors such as TBP bind to DNA. [2]

1) The TBP has a DNA-binding site* which is complementary in shape and charge to the
TATA sequence;
2) which comprises a specific sequence of bases and hence has a specific shape;

(b) Explain why the unwinding of the double helix of DNA promotes transcription.[2]

1) The unwinding of the double helix of DNA exposes the template for complementary
base pairing* with free ribonucleotides;
2) RNA polymerase can then catalyse the formation of phosphodiester bonds* between
the free ribonucleotides, forming the mRNA;

(c) Explain why such processing of pre-mRNA molecules is necessary. [3]

1) Splicing* of pre-mRNA involves cutting out introns and joining exons to form a mature
mRNA*;
2) introns which are non-coding regions are removed;
3) The mature mRNA comprises exons which code for the sequence of amino acids in a
protein;

[Total: 7]
Raffles Institution Nov 2012 (H2 Biology) Paper 2 (for 9744 syllabus)

N12P2Q5

(a) Explain the term epistasis in this context.[3]

1. Epistasis is a form of gene interaction in which a gene at one locus alters the phenotypic
expression of a gene at a second locus;
2. e.g. allele A of one gene codes for a functional enzyme A, which converts the colourless
precursor into a colourless intermediate. Allele B of another gene codes for enzyme B,
which converts the colourless intermediate into the purple pigment;
3. Genotype aa masks genotype BB and Bb. Thus lack of functional enzyme A in pea plant
will result in white flowers despite presence of functional enzyme B;

(b) Use the symbols A, a and B, b to draw a genetic diagram to explain the results shown in the
F2 generation. [4]

1) Parental genotype aaBB X AAbb

Parental phenotype white flower white flower

2) F1 genotype All purple

F1 phenotype AaBb

3) Gametes AB aB Ab ab x AB aB Ab ab

4) punnett square

AB aB Ab ab

AB AABB, purple AaBB, purple AABb, purple AaBb, purple

AaBB, purple aaBB, white AaBb, purple aaBb, white

aB

AABb, purple AaBb, purple AAbb, white Aabb, white

Ab

AaBb, purple aaBb, white Aabb, white aabb, white

ab

5) F2 genotypic ratio
9 A_B_ : 3 A_bb : 3 aaB_ : 1 aabb

F2 phenotypic ratio 9 purple: 7 White

[4]
Raffles Institution Nov 2012 (H2 Biology) Paper 2 (for 9744 syllabus)

(c) Explain how the chi-squared calculated value of 1.60 supports the statement that epistasis
is the correct explanation of these results. [3]

1) The chi-squared calculated value of 1.60 means 0.1 < p < 0.5;
2) At significant level of 5%, since p is greater than 0.05, we do not reject the null
hypothesis that the expected results are similar to the observed results;
3) Thus, there is no significant difference between the numbers of expected and observed
phenotypes and hence the deviation is due to chance;
4) The F2 ratio is close to 9 purple : 7 white flower, a modified 9:3:3:1 ratio which indicates
epistasis;

(d) Suggest why pea plants are good experimental organisms for carrying out such crosses. [2]

1) Pea plants grow fast so we don’t have to wait long to study the different characteristics
of the plants appearing in many generations;
2) Pea plants have many observable traits with contrasting forms e.g. round seed vs
wrinkles seeds;
3) Pea plants produce many offspring in one cross;
4) There is ease in manipulating pollination – can do cross or self-pollination;
[Total: 12]

N12P2Q5

(a) Explain why there is a smaller yield of ATP per molecule of glucose from anaerobic
respiration than aerobic respiration. [4]

1) Only 2 ATP* is generated per glucose molecule during anaerobic respiration as

compared to 36-38 ATP* for aerobic respiration.;
2) From anaerobic respiration without oxygen*, which is the final electron acceptor*,
the electron flow through electron transport chain (ETC) is disrupted, thus oxidative
phosphorylation* cannot occur ;
3) In aerobic respiration, glucose is completely oxidised to carbon dioxide, whereas in
anaerobic respiration, incomplete oxidation occurs to produce pyruvate;
4) glycolysis of each glucose molecule produces 2 ATP through substrate level
phosphorylation under anaerobic conditions;
5) Oxidation of NADH in the ETC during aerobic respiration yields the 34-36 ATP while
oxidation of NADH in anaerobic respiration does not yields any additional ATP;

(b) Using your knowledge of ATP yields, calculate an estimate of how many times faster
glycolysis must be under anaerobic conditions when compared to aerobic conditions in
order to produce the same number of ATP molecules in the same time.

Show your working clearly below. [3]

1) Glycolysis which can occur under ana
2) erobic condition produces 2 ATP net
3) Oxidative phosphorylation produces 34-36 ATP
4) Glycolysis must occur 34/2 or 36/2 times the speed of oxidative phosphorylation in
order to produce the same number of ATP
Number of times faste _____17-18 times __________
Raffles Institution Nov 2012 (H2 Biology) Paper 2 (for 9744 syllabus)

(c) Describe and explain the effect on PFK activity of increasing ATP concentration.[4]

1. High levels of ATP slows down / inhibits PFK activity (as the graph is below the graph
represented by low ATP);
2. This inhibition is most pronounced at lower substrate concentrations
3. this inhibition is overcome by high substrate concentrations
4. ATP binds to allosteric site* of PFK
5. altering its active site conformation such that the substrate (F6P) cannot bind thus
lowering the PFK activity.

(d) Explain the meaning of the term facilitated diffusion. [3]

1) Facilitated diffusion is a process of passive transport where no energy is required

2) And it occurs down a concentration gradient
3) And takes place via protein channels which allows specific molecules to move across
the membrane
An example is the movement of charged or polar molecules down concentration gradient via
the transmembranal transport protein that provide a hydrophilic pore across the membrane
[Total: 14]

N12P2Q7

(a) Explain the ways in which islands favour the formation of new species. [6]

1. Islands are geographically isolated* as they are surrounded by water that acts as a
physical barrier preventing interbreeding. This results in the disruption of gene flow*;
2. The islands due to their differing habitats / environments, present many niches for the
species to fill – idea of adaptive radiation;
3. e.g. of differences. Soil type – sandy or clayey, availability of water, availability of shade,
plant types;
4. Thus the (common) ancestral species on different islands were exposed to different
selection pressures* and natural selection act on them;
5. There exist variation in the population and those with advantageous characteristics/best
adapted to the local conditions are more likely to survive, reproduce and pass on their
alleles to the next generation;
6. As the different populations evolve independently from each other, their allele frequencies
change and they accumulate different genetic mutations over time;
7. Allele frequencies change due to natural selection and genetic drift/founders effect (which
are random events that are due to chance);
8. Over hundreds and thousands of generations, across long periods of time, accumulation of
genetic differences led to each population on different islands becomes reproductively
isolated*;
9. Eventually they can no longer interbreed* to produce viable, fertile* offspring and hence
new species are formed through allopatric speciation;
Raffles Institution Nov 2012 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Suggest advantages of using mitochondrial DNA. [2]

1. There is no recombination/crossing over in mitochondrial DNA from parent to offspring

(idea of no recombination) unlike nuclear DNA, hence changes in DNA sequence is
SOLELY due to the accumulation of mutations over time so it can reliably be linked to
the molecular clock;
2. Faster mutation rate (key idea) compared to nuclear DNA and hence it is useful for
comparing individuals within a species or species that are closely related* as you
require discernible differences between the DNA of organisms being compared;

(c) Describe the advantages of using nucleotide data such as this in classifying
organisms.[3]

1. The nucleotide data are objective. Molecular character states are unambiguous as A, C,
G and T are easily recognisable and cannot be confused.
2. The nucleotide data are quantitative. Molecular data are easily converted to numerical
form and hence are amenable to mathematical and statistical analysis and hence
computation. The degree of relatedness can be inferred and quantified by calculating the
nucleotide differences between species.
3. The nucleotide data can be used to compare species which are morphologically
indistinguishable due to convergent evolution or are very closely related.
4. Remotely related organisms such as bacteria, humans and sunflower can also be
compared because they share some proteins such as cytochrome c.

(d) Outline two of the functions of these protein complexes. [2]

1. One of the protein complexes is ATP synthase *which is an enzyme which couples the
phosphorylatione of ADP to ATP to the movement of H+ down its concentration
gradient via the process of chemiosmosis;
2. Other protein complexes are electron carriers* that alternate between reduced and
oxidized states as they accept and donate electrons respectively. They help to facilitate
the flow of electrons in the electron transport chain found in the inner membrane of
mitochondria used for oxidative phosphorylation.
[Total: 13]
Raffles Institution Nov 2012 (H2 Biology) Paper 2 (for 9744 syllabus)

N12P2Q8

8. (a) Outline the molecular structure of phospholipids in relation to their function in cell
membranes. [7]
Structure Function

Two non-polar*, hydrophobic* Major component of the cell

hydrocarbon tails/chains* membrane
associated with a glycerol* backbone; that can form a phospholipid
bilayer* (if not mentioned under
The remaining site is joined to a charged* structure);
(reject polar!) hydrophilic* phosphate* in an aqueous environment that acts
group making it amphipathic; as a barrier* to polar and
charged molecules/ions*;
phospholipid The hydrophilic heads face outwards and
interact with the aqueous environment of They control the permeability of the
the cell interior or exterior; whilst the membrane /acts as a selectively
hydrophobic tails face inwards, away from permeable membrane because the
the water ; hydrophobic core
giving rise to a cell membrane that is made region is only permeable to small
up of a phospholipid bilayer * with a hydrophobic solutes ;
hydrophobic core*;

(b) Explain the role of the Golgi body and its link to the rough endoplasmic reticulum.

1. rER has ribosomes* that stud the outer surface of the membrane that are involved in
translation* of mRNA into proteins
2. Proteins formed enter the cisternal space, where they fold into their specific
conformation and undergo modification
3. Vesicles carrying the proteins then bud off from the rER to the cis face of Golgi
apparatus for further modification;
4. Consists of a stack of flattened, membrane-bound sacs called cisternae*, together with
associated Golgi vesicles;
5. Vesicles* from the rough ER carrying proteins will fuse* with the cis face of the Golgi
apparatus, releasing their contents into the cisternal space;
6. Where they will be further modified by glycosylation* e.g. oligosaccharide on
glycoproteins can be modified to give a variety of glycoprotein products;
7. Golgi vesicles bud off from the trans face of the Golgi apparatus, carrying the finished
products;
8. Products of modification are sorted and transported to various parts of the cell, e.g. the
plasma membrane for insertion into the membrane or secretion through exocytosis*;
9. Also involved in the formation of lysosomes*;
Raffles Institution Nov 2012 (H2 Biology) Paper 2 (for 9744 syllabus)

(c) Describe the structure of ribosomes. [6]

1.
Ribosomes are complexes of ribosomal RNA* and proteins*.
2.
Each ribosome consists of a small subunit and
3.
a large subunit
4.
Prokaryotes have 70S ribosomes, each consisting of a small (30S) and a large (50S)
subunit
5. Eukaryotes have 80S ribosomes, each consisting of a small (40S) and a large (60S)
subunit
6. The ribosomes have a peptidyl-tRNA binding site (P site) and amino-acyl tRNA binding
site (A Site) and exit siteE(exit);
7. Part of an rRNA molecule in the small subunit has an mRNA binding site;
8. Part of an rRNA molecule in the large subunit has the ability to catalyse peptide bond
formation;
Where S* = Svedberg units (unit for sedimentation rate)

[Total: 20]

N12P2Q9

9.
(a) Outline the light dependent reactions of photosynthesis [7]

1. Light energy is absorbed* by accessory pigments molecules in light harvesting

complex of Photosystems (PS) I and II*;
2. The electrons in these pigment molecules become excited and, when returning to their
ground states, pass on the released energy to the next pigment molecule and excite
the electrons present in them, as a result. This resonance transfer of energy occurs
until it reaches one of the two special chlorophyll a molecules (P700 in PS I & P680 in
PS II) in the reaction centre;
3. When the special chlorophyll a molecule absorbs the energy, an excited electron is
displaced (Chl a chl a+ +e-) leaving an electron hole in PS I and II.
4. This electron is captured by a primary electron acceptor molecule in the reaction
centre;
5. The excited electrons flow down a chain of carriers of increasing electronegativity along
the electron transport chain, which links PS II to PS I.
6. The energy lost from the redox reactions which occur along this electron transport
chain is used to actively pump protons across the thylakoid membranes into the
thylakoid space, resulting in a proton/H+ gradient* forming across the thylakoid
membrane;
7. As proton/H+ diffuses down its concentration gradient back into the stroma via the ATP
synthase*, ADP is phosphorylated to ATP. This process is called chemiosmosis;
8. The splitting of water by an enzyme (H2O 2e─ + 2H+ + ½O2) releases protons/H+ and
+
H diffuse from thylakoid space to stroma through ATP synthase*;
9. The electron produced from the splitting of water, fills the electron hole in PSII and the
electron from PSII fills the electron hole in PSI
10. Electrons from PS I are passed down an electron transport chain and are finally
transferred to NADP which combines together with H+ to form reduced NADP in
stroma. This reduction is catalyzed by NADP reductase;
Raffles Institution Nov 2012 (H2 Biology) Paper 2 (for 9744 syllabus)

11. NADP is the final electron acceptor* in the non-cyclic light dependent reaction*;
12. Reduced NADP is used in the Calvin cycle* in the stroma of the chloroplast;

(b) Explain the role of membranes in the chloroplast. [7]

1. The thylakoid membrane maintains the specific positions of the protein molecules and
chlorophyll molecules making up the photosystems
2. as well as the positions of the molecules that make up the electron transport chain
3. to ensure transfer of energy and the flow of electrons between the photosystems and
electron transport chain;
4. The thylakoid membrane also contains ATP synthase* and maintains its
position/orientation so that
5. ATP can be produced as protons flow down their gradient (chemiosmosis) from
thylakoid space to stroma;
6. Maintains proton gradient* for ATP synthesis since the hydrophobic core* of the
membrane is impermeable to protons and this is essential for chemiosmosis;
7. The stacks of thylakoid membranes also provides a large surface area to embed many
chlorophyll molecules (hydrocarbon tail of chlorophyll interacts with hydrophobic core of
thylakoid membrane) for light absorption* and for the light energy excite the electrons
of the chlorophyll molecules;

(c) Describe the effect of increasing light intensity on the rate of photosynthesis. [6]

1. At P, i.e. low light intensities, the rate of photosynthesis increases linearly with
increasing light intensity
2. as light intensity is a limiting factor*.
3. At Q: the rate of increase of photosynthesis starts to fall as light intensity is not the only
limiting factor.
4. Some other factor such as CO2 concentration, is also limiting.
5. At R: light intensity is no longer limiting. Even when light intensity is increased, there is
no increase in the rate of reaction. Some other factor is limiting.
6. At S: Light saturation point: Light intensity beyond which an increase in light
intensity will not increase the rate of photosynthesis

[Total: 20]
Raffles Institution Nov 2012 (H2 Biology) Paper 3 (for 9744 syllabus)

Nov 2012 H2 Bio Paper 3

N12P3Q1

(a)– (c) (OUT OF SYLLABUS)

(d)
(i) Describe what occurs at stage Y. [2]

1. Primer annealing step which occurs at 64 C in the presence of a large excess of the
DNA primers* ;
2. allowing their specific attachment to the single stranded complementary DNA template
by complementary base pairing* to the regions flanking the target sequence to be
amplified.;

(ii) If the automated process had been stuck at stage X for two hours and then allowed
to slowly cool down to room temperature, describe what you would expect to find in
the PCR mixture. [3]

1. There would be Taq polymerase;

2. Deoxyribonucleotides;
3. forward and reverse primers;
4. double the amount of double stranded DNA (as one cycle had occurred);

(e) Describe what is happening at A and B. [2]

A – denaturation* of DNA* by breaking of hydrogen bonds* between complementary* base

pairs resulting in single stranded DNA formed
B – nucleic acid hybridisation occurs where a probe which is a short single-stranded DNA
molecule forms hydrogen bonds via complementary base pairing* with complementary
sections of the single stranded DNA
[Total: 15]

N12P3Q2

(a) State the potency of the stem cells found in the bone marrow. [1]
Multipotent

(b) Suggest and explain why all the cells were removed from the donor trachea.[3]
1. To avoid stimulation of patient’s immune system
2. because donor cells are recognised as foreign
3. which will eventually cause trachea to be destroyed by patient’s immune system (idea)
4. To make space for donor cells to be replaced by new cells differentiated from patient’s
bone marrow stem cells.
Raffles Institution Nov 2012 (H2 Biology) Paper 3 (for 9744 syllabus)

(c) Suggest and explain why the stem cells needed to be treated with chemicals to stimulate
proliferation. [3]
1. These chemicals stimulate mitosis* produce identical daughter cells capable of
2. self-renewal* producing daughter cells that possess same developmental and
differentiation potential / potency as parent cell
3. Stem cells are able to differentiate* to form specialised cells upon receiving appropriate
molecular signals / chemical signals which can come in the form of chemicals used to
treat them
4. These chemicals cause changes in pattern of gene expression / switching on-off
different sets of genes that confer a particular function, resulting in specialisation.

(d) Suggest how stem cells can continue to divide mitotically, whilst other cells can only do this
for a set number of divisions. [3]
1. Each round of DNA replication is accompanied by DNA replication which involves
shortening of telomeres* at ends of chromosomes
2. When telomeres are shortened to a critical length, cells are no longer able to divide
(does it stop dividing? check non functional, apotosis?)
Suggested answer:
When telomeres are shortened to a critical length, cells stop dividing / divide a limited
number of times /undergo programme cell death/ triggered pathway to apoptosis,
3. Stem cells express telomerase gene to produce telomerase* that lengthens telomeres
thus allowing them to proliferate continuously.
3. But in other cells, expression of telomerase gene is switched off.

[Total: 10]

N12P3Q3 (OUT OF SYLLABUS)

N12P3Q4
Planning
1. Aim : To investigate the effect of concentration of sucrose solution on the rate of respiration
in yeast cells.

2. Theory
(Main theory)
Sucrose is a dissacharide that can be hydrolysed by yeast cells into glucose and
fructose. The monosaccharide glucose, used for glycolysis to produce 2 pyruvate
which then undergoes the link reactions to produces 2 CO2 and 2 acetyl CoA. The 2
acetyl CoA enters the Krebs cycle and produces 4 molecules of CO2.

As the concentration of sucrose increases, more glucose will be produced. The

increase in the concentration of glucose will result in higher rate of glycolysis, linked
reaction and Krebs thus resulting in greater rate of production of CO2.

The rate of production of CO2 can be measured by measuring the mass of CO2
produced per unit time. The mass of CO2 produced can be measured by bubbling
CO2 through barium hydroxide containing phenolphthalein indicator for 5 minutes.
The resultant solution is then titrated using 0.1 moldm-3 HCl. The volume of 0.1
Raffles Institution Nov 2012 (H2 Biology) Paper 3 (for 9744 syllabus)

moldm-3 HCl (x) required to neutralise the barium hydroxide causing the
phenolphthalein to change from pink to colourless is then recorded.

To calculate the mass of CO2 produced, use the following formula

Mass of CO2 produced /mg = (5 – x) x 2.2 mg

Increasing the concentration of sucrose solution will result in a higher rate of CO2
production and thus decreasing the volume of HCl required to change the
phenolphthalein from pink to colourless.

3. Procedure (PAN CR)

a) [P] Pilot test*
Conduct a pilot experiment to determine suitability of apparatus, suitability of range of
independent variable (e.g. concentration of sucrose solutions), optimum conditions, and
amount of materials used (volume and concentration of yeast suspensions used)

b) Annotated diagram

c) Variables
[DV] Dependent variable AND description of how it is measured : Volume of 0.1
moldm-3 HCl need to turn phenolphthalein from pink to colourless. The volume is measured
using a burette.
[IV] Independent variable (at least 5 concentrations at equal interval) AND
description of how it is controlled : 0.0, 0.2, 0.4, 0.6, 0.8 and 10 sucrose moldm-3
solution by diluting 10% sucrose solution using the dilution table below :

Concentration of sucrose Volume of 1.0 moldm-3 Volume of distilled water

solution / moldm-3 sucrose solutions used used /cm3
/cm3
0.0 0.0 10.0
0.2 2.0 8.0
0.4 4.0 6.0
0.6 6.0 4.0
0.8 8.0 2.0
1.0 10.0 0.0

[CV] Controlled variable and description of how it is controlled:

Concentration of yeast suspension used
Temperature
Volume of yeast suspension used
Volume of yeast suspension used
Concentration of HCl used
Volume of barium hydroxide used
Concentration of barium hydroxide used
Concentration of phenolphthalein used
Volume of phenolphthalein used
Raffles Institution Nov 2012 (H2 Biology) Paper 3 (for 9744 syllabus)

d) Numbered steps
1. Prepare a 30oC water bath by mixing hot and cold water in a 500cm3 beaker. Use a
thermometer to measure the temperature and adjust the temperature to 30oC by mixing
hot and cold water.

2. Using a 5.0 cm3 syringe, measure and dispense 5.0 cm3 of active yeast suspension into
5 separate test tubes and place the test tubes in the 30oC water bath.

3. Prepare various concentrations of sucrose solutions by measuring the following

volumes of sucrose solutions and distilled water using a 10 cm3 syringe and dispensing
them into respectively labeled test tubes.
Concentration of sucrose Volume of 1.0 moldm-3 Volume of distilled water
-3
solution / moldm sucrose solutions used used /cm3
3
/cm
0.0 0.0 10.0
0.2 2.0 8.0
0.4 4.0 6.0
0.6 6.0 4.0
0.8 8.0 2.0
1.0 10.0 0.0

4. Shake to mix the sucrose solutions well and place the solution in the 30oC water bath.

5. Using a stopwatch, time for 5 minutes to allow the yeast suspension and sucrose
solution to reach 37oC. This is the equilibration time.

6. Using a 10cm3 syringe, measure and dispense 10 cm3 of 0.025 moldm-3 barium
hydroxide solution and 1 cm3 of 1% phenolphthalein into 6 test tubes. Shake to mix.

7. Pour the active yeast suspension into the test tube containing 0.2 moldm-3 sucrose
solution. Mix using a glass rod and set up using delivery tube and 0.025 moldm-3 barium
hydroxide solution as shown in the diagram below

8. Start the stop watch and time for 10 minutes. After 10 minutes, remove the test tube
containing mixture of barium hydroxide solution and phenolphthalein.

9. Draw 10cm3 of 0.1 moldm-3 hydrochloric acid into a 10cm3 syringe. Add the hydrochloric
acid dropwise shaking after every drop until the phenolphthalein turn from pink to
colourless. Record the volume of 0.1 moldm-3 hydrochloric acid used (x). This is the
dependent variable.

10. Calculate the mass of CO2 produced in 10 minutes using the following formula.
Mass of CO2 produced /mg = (5 – x) x 2.2 mg

11. Repeat step 7 to 10 using 0.4, 0.6, 0.8, 1.0 moldm-3 sucrose solution. This is the
independent variable.

12. Repeat Step 7 to 11 twice to serve as replicates to check for anomalous results

13. Repeat the entire experiment twice to check for reproducibility of results.
Raffles Institution Nov 2012 (H2 Biology) Paper 3 (for 9744 syllabus)

e) [C] Control
Keeping all other variable such as volumes, concentrations of yeast suspensions and
barium hydroxide solutions and temperature constant, use distilled water instead of
sucrose solution. This is to show that any change in the mass of CO2 produced is due to
the sucrose solutions.

4. Data Recording and Processing

Record and process data as follows:

Concentration Volume of 0.1 moldm-3 hydrochloric acid used Mass of CO2

of sucrose to turn phenolphthalein from pink to colourless produced in 10
solution / /cm3 mins / mg
-3
moldm Replicate Replicate Replicate Average
0.0 1 2 3
0.2
0.4
0.6
0.8
1.0
[G] Graph showing the effects concentration of sucrose solution on the rate of respiration
in yeast cells.

Mass of CO2
produced in 10
mins / mg

Concentration of sucrose
solution / moldm-3

Risks & Precautions

a. The barium hydroxide and hydrochloric acid solutions are corrosive when it comes
in contact with skin and eyes. Wear protective gloves and goggles when handling
these solutions.
b. The activated yeast suspension may be a biohazard and may cause infection
when it contact with skin and eyes. Wear protective gloves and goggles when
handling these solutions.
c. The hot water used to make water bath may cause scalding. Wear mitten when
handling the hot water.

N12P3Q5 (OUT OF SYLLABUS)

Raffles Institution Nov 2013 (H2 Biology) Paper 2 (for 9744 syllabus)

Nov 2013 H2 Bio Paper 2

N13P2Q1

(a) Describe the processes that occur in the Golgi body. [3]
1. Glycosylation* where oligosaccharides/short sugar chains are added to proteins
and lipids to form glycoproteins and glycolipids respectively;
2. Modification of existing glycoproteins and glycolipids made in the endoplasmic
reticulum by modifying the existing oligosaccharide / cleaving sugar molecule(s)
from the oligosaccharide;
3. In plant cells, Golgi body is the site for synthesis of polysaccharides such as pectin
(a component in plant cell wall) and then transported in vesicles to the cell
membrane;
4. Sorting* of proteins into different kinds of vesicles to target the proteins to different
parts of the cell or for secretion;

(b) Describe the role of vesicles that fuse with the forming face of the Golgi body. [3]
1. Vesicles that bud off from rough endoplasmic reticulum (RER) carry proteins;
2. and bud off from smooth endoplasmic reticulum (SER) carry lipids;
3. These vesicles are targeted to fuse to cis-face of Golgi by microtubules of the
cytoskeleton/proteins;

(c) Outline two roles for the vesicles that are formed at the maturing face of the Golgi body. [4]
1. Lysosome / lysosomal vesicles;
2. fuses with vesicles/vacuoles formed by endocytosis to digest the contents (e.g.
foreign particles) within the vesicles/vacuoles; (pt 1 & 2 = 2 marks)
3. Exocytic vesicles;
4. fuses with plasma membrane to release enzymes outside of cell by exocytosis to
breakdown extracellular content / fuses with plasma membrane to release contents
outside of cell by exocytosis; (pt 3 & 4 = 2 marks)
5. Secretory vesicles;
6. fuses with plasma membrane to release contents upon receiving signal; (pt 5 & 6 =
2 marks)

Any 2 pairs of points

N13P2Q2

(a) State what happens if a cell loses control of the cell cycle. [1]
1. Uncontrolled cell proliferation;

(b) With reference to Fig. 2.1, suggest how the dysregulation of checkpoints of cell division
may occur. [3]
1. Point mutation of S-cyclin gene to cause change in amino acid sequence of S-
cyclin protein, thus S-cyclin is resistant to degradation / Point mutation of M-cyclin
gene to cause change in amino acid sequence of M-cyclin protein, thus M-cyclin is
resistant to degradation;
2. Prolonged activation of M-CDK and S-CDK to cause more rounds of cell division
thus excessive cell proliferation;
3. Gene amplification of CDK gene to increase number of copies of gene resulting in
increase amount of CDK protein;
4. Increase number of binding sites for S-cyclin/M-cyclin to form increase amount of
active CDK;
Raffles Institution Nov 2013 (H2 Biology) Paper 2 (for 9744 syllabus) 2017

(c) (i) Name one causative agent of cancer. [1]

1. Viruses e.g. HPV, avian sarcoma virus;
2. Ultraviolet light / Ionising radiation (mention of radiation along is insufficient);
3. Carcinogens e.g. tar in cigarette smoke, asbestos, benzene, formaldehyde,
ethidium bromide;
Any one

(ii) Outline the development of cancer including the effects of this causative agent. [5]
1. The causative agent increase chances of DNA damage and mutations in the
genes which control regulatory checkpoints of the cell cycle in a single cell;
2. Loss-of-function mutation of tumor suppressor genes will result in inability to inhibit
cell cycle, repair damaged DNA and promote apoptosis;
3. Gain-in-function mutation of proto-oncogenes to form oncogenes will result in
overexpression of proteins/growth factors OR
production of hyperactive/degradation resistant proteins/growth factors;
4. leading to excessive cell proliferation/division to form tumour;
5. Loss of contact inhibition enables cells to grow into a tumour;
6. Activation of genes coding for telomerase so that cells can divide indefinitely;
7. Angiogenesis must occur within the tumour so that the blood vessels formed can
transport oxygen and nutrients for its growth;
8. Resulting in the formation of a maglinant tumour capable of metastasizing to other
parts of body to form secondary tumours;

N13P2Q3
Fig.3.1 shows the mechanism of DNA replication originally proposed by Watson and Crick.

(a) State the name given to this mechanism of DNA replication. [1]
Semi-conservative DNA replication*

(b) Describe how these results provided evidence for Watson and Crick’s proposed mechanism.
[3]
1. In generation 0 only the heavy 15N-15N DNA molecules were present which appeared as
the lowest band in the caesium chloride solution.
2. During semi-conservative replication, the original 15N-15N DNA strands unzipped* and
served as templates* for the formation of the new strands. Since only 14N DNA was
present in the medium, the resulting DNA molecules in generation 1 were hybrid* DNA
molecules where each molecule was made up of one original 15N strand and one new
14
N strand. This hybrid DNA molecule thus appeared as the intermediate band in the
caesium chloride solution.
3. Each hybrid DNA molecule from generation 1 unzipped and were used as templates for
DNA replication. Hence 50% of the DNA in generation 2 was made up of hybrid DNA
(i.e. 15N-14N) which appeared as the intermediate band and 50% was made up of light
DNA (i.e. 14N-14N) which appeared as the uppermost band in the caesium chloride
solution.

Thus the appearance of the various bands at the various generations is consistent with
semi-conservative replication.
Raffles Institution Nov 2013 (H2 Biology) Paper 2 (for 9744 syllabus) 2017

(c) List three ways in which transcription is different from DNA replication.[3]

Feature DNA replication Transcription

Template In replication, both DNA strands serve In transcription, only one
as template. strand acts as template.
Product 2 double-stranded DNA molecules are 1 single stranded RNA
synthesised. molecule is synthesised.
Polymerase DNA polymerase* catalyses the DNA-dependent RNA
involved reaction. polymerase* catalyses the
reaction.
Process The process is associated with cell and The process is associated
associated with nuclear division. with protein synthesis.
Base that pairs Thymine is used during the process. Uracil is used during the
with Adenine process.
Process Both DNA strands are used as One DNA strand is used as
template in the synthesis of DNA. template for the synthesis of
mRNA.
Nucleotides Deoxyribonucleotides and Ribonucleotides are used.
used ribonucleotides (for RNA primer) are
used
Comments: The main differences between transcription and replication were well known to
most candidates and many provided full responses.

(d) Explain how the information to synthesise polypeptides is coded for by DNA.[3]

1. A specific sequence of nucleotides on a template DNA strand is transcribed by RNA

polymerase to form mRNA by complementary base pairing*;
2. The mRNA contains triplet base codes known as codons* that each codes for a
specific amino acid;
3. When the mRNA containing a specific sequence of codons which code for a specific
amino acid sequence,is translated at ribosomes, polypeptides are formed.

N13P2Q4
Fig.4.1 shows an outline of the first three stages of aerobic respiration

(a) Name the stages A, B and C and state precisely where in a eukaryotic cell these stages
occur.[3]
A: Glycolysis occurs in the cytosol
B: Link reaction occurs in the matrix of the mitochondrion
C: Krebs cycle occurs in the matrix of the mitochondrion
Comments: Candidates demonstrated sound knowledge and understanding of aerobic
respiration. Most candidates were able to identify where the stages occurred. Some
candidates were not precise in their responses.

(b) For each glucose molecule, state the total number of ATP formed as a result of stages A
and C, including any ATP produced through oxidative phosphorylation of the products.[2]
A: 7 (2 ATP + 2NADH where each NADH produces 2.5ATP = 2 +5 =7)
C: 20 (2 ATP + 6NADH where each NADH produces 2.5ATP + 2 FADH where each
FADH produced 1.5ATP = 2 +15 + 3 =20)

Also accept: A:8 and C:24 (as students may use 1 NADH produced 3ATP and 1 FADH
produced 2 ATP)
Raffles Institution Nov 2013 (H2 Biology) Paper 2 (for 9744 syllabus) 2017

Oxidative phosphorylation regenerates FAD and NAD from the reduced FAD and reduced NAD.
(c) Outline how this results in the production of ATP.[4]
1. Reduced NAD and FAD transfer their high energy electrons to the electron acceptors of
the electron transport chain, and get re-oxidised in the process;
2. As electrons are passed down electron carriers of increasing electronegativity, the
energy release is coupled to the pumping of H+ into the intermembrane space to
generate a proton motive force/proton gradient;
3. As H+ ions flow through the ATP synthase back into the matrix down the proton gradient,
ATP is produced from ADP and inorganic phosphate via chemiosmosis;
4. Reoxidation of reduced NAD and FAD allows the regeneration of the coenzyme NAD
and FAD, allows them to pick up more protons and electrons from Krebs cycle, link
reaction and glycolysis, so that these reactions can continue;

N13P2Q5

(a) Suggest why the IA and IB alleles are dominant over the i allele. [3]
1. The expression of the dominant IA or IB allele masks the effect of the recessive i allele;
2. The presence of IA or IB allele leads to the production of A antigens/glycoproteins or B
antigens/glycoproteins respectively on the surface of the red blood cells;
3. Therefore heterozygous individuals, with IAi or IBi genotypes, have A antigens or B
antigens, respectively, on their red blood cells. This suggests that IA and IB alleles are
dominant over the i allele;

(b)
(i) The ABO blood type gene is located on chromosome nine. Suggest where the Rhesus
(Rh) factor gene may be found. Explain the reason for your answer. [2]
1. Not found on chromosome 9
OR
Found on another chromosome apart from chromosome 9;

2. As the alleles coding for the Rhesus factor are inherited independently, they are
not linked to the alleles of the ABO blood types. Thus, they are not found on the
same chromosome;
 Raffles Institution Nov 2013 (H2 Biology) Paper 2 (for 9744 syllabus) 2017

(ii) A type A Rhesus positive mother and a type B Rhesus positive father have a type O
Rhesus negative child.

Draw a genetic diagram in the space below to show how this is occurred.

Use the symbols given above and show all the possible genotypes and phenotypes for
the offspring of these parents.

Parental phenotype: Type A, Rhesus positive x Type B, Rhesus positive

Parental genotype: IAiRh+Rh- x IBiRh+Rh-
Meiosis
Gametes:
IARh+ IARh- iRh+ iRh- x IBRh+ IBRh- iRh+ iRh-
Random fusion of gametes:

IARh+ IARh- iRh+ iRh-

IAIBRh+Rh+ IAIBRh+Rh- IBiRh+Rh+ IBiRh+Rh-
B +
I Rh
Type AB, Rhesus Type AB, Rhesus Type B, Rhesus Type B, Rhesus
positive positive positive positive
IAIBRh+Rh- IAIBRh-Rh- IBiRh+Rh- IBiRh-Rh-

IBRh- Type AB, Rhesus Type AB, Rhesus Type B, Rhesus Type B, Rhesus
positive negative positive negative
IAiRh+Rh+ IAiRh+Rh- iiRh+Rh+ iiRh+Rh-
+
iRh
Type A, Rhesus Type A, Rhesus Type O, Rhesus Type O, Rhesus
positive positive positive positive
IAiRh+Rh- IAiRh-Rh- iiRh+Rh- iiRh-Rh-
iRh-
Type A, Rhesus Type A, Rhesus Type O, Rhesus Type O, Rhesus
positive negative positive negative

Offspring genotype:
AB + + AB + - AB - - B + + B + - B - - A + + A + - A - - + + + - - -
I I Rh Rh , I I Rh Rh , I I Rh Rh , I iRh Rh , I iRh Rh , I iRh Rh , I iRh Rh , I iRh Rh , I iRh Rh , iiRh Rh , iiRh Rh ,iiRh Rh

Type Type B, Type B, Type A, Type A, Type O, Type O,

Type AB,
Phenotype AB, Rhesus Rhesus Rhesus Rhesus Rhesus
Rhesus Rhesus
Rhesus positive negative positive negative positive negative
negative
positive

Offspring phenotypic ratio:

Type AB, Rhesus positive : Type AB, Rhesus negative: Type B, Rhesus positive: Type B, Rhesus negative: Type A,
Rhesus positive: Type A, Rhesus negative: Type O, Rhesus positive: Type O, Rhesus negative

3:1:3:1:3:1:3:1
Therefore, there is a 1 in 16 chance of the child having a Type O, Rhesus negative phenotype

[5]
Raffles Institution Nov 2013 (H2 Biology) Paper 2 (for 9744 syllabus) 2017

N13P2Q6
Fig. 6.1 represents the behaviour of one pair of chromosomes during meiosis.

(a) Name the structures A, B and C. [3]

A: Homologous chromosomes/ Bivalents
B: Centromere
C: Sister chromatid

(b) (i) Explain how structure C is similar to D. [2]

1) C & D are non-identical sister chromatids of homologous chromosomes;
2) They have the same sequence of genes (e.g. genes coding of X,Y and Z) at the
same loci;
3) They are also joined to their own identical sister chromatid at the same
centromere position;

(ii) Explain how the structure of C is different from structure D. [2]

1) Structures C & D carry different alleles at the same gene loci (e.g. C carries
dominant allele Y while D carries recessive allele y);
2) This is due to each homologue being inherited from each of the parents (i.e.
maternal and paternal chromosomes);

(c) Describe what is occurring during stage 2 and explain its effects on the products of meiosis.
[4]
1) Following synapsis to form bivalents (stage 1), crossing over* occurs between non-
identical sister chromatids of homologous chromosomes;
2) During crossing over, bivalents are seen as tetrads;
3) The sites at which non-sister chromatids of homologous chromosomes break and rejoin
with each other are known as chiasmata;
4) This process leads to different allelic combinations being found on the sister chromatids,
which eventually separate, following meiosis II, to become individual chromosomes in
gametes (stage 3);
5) Thus, this increases genetic variation within a population. Allelic combinations which
confer advantageous phenotypes to individuals are selected for due to natural selection
and increase in frequency over time;

N13P2Q7

(a) Describe the properties of the phospholipid bilayer and the aquaporin channels in relation to
the movement of water across the cell surface membrane. [4]
1. Phospholipids have hydrophilic phosphate heads* which face outwards and interact
with the aqueous environment of the cell interior or exterior;
2. whilst the hydrophobic hydrocarbon tails* face inwards, away from the water giving
rise to a lipid bilayer;
3. Phospholipids have a hydrophobic core region that prevents movement of polar water
molecules across the bilayer;
4. Aquaporin channels are protein channels which are made up of amino acids with
hydrophobic R groups that are able to interact with the hydrophobic core region of the
phospholipid bilayer;
5. The interior of the channel is made up of amino acids with hydrophilic R groups which
allow movement of the polar water molecules through the channel pore;
Raffles Institution Nov 2013 (H2 Biology) Paper 2 (for 9744 syllabus) 2017

(b) Explain what has happened to the treated cells after 3 minutes. [4]
1. The treated cell appeared to increase in size and eventually lyse after 3 minutes as
compared to the control cell which did not change in size within the same duration;
2. The genes coding for the aquaporin channels injected into the treated cells were
expressed;
3. resulting in more aquaporin channels in the cell surface membrane compared to the
control cells;
4. More water molecules were able to enter the treated cells by osmosis causing it to
increase in volume and eventually rupture;

(c) Outline the differences between osmosis and facilitated diffusion. [2]
1. Osmosis involves the diffusion of water molecules from a region of higher water potential
(less negative water potential) to a region of lower water potential (more negative water
potential). Facilitated diffusion involves the diffusion of polar (or charged) molecules or
ions, unable to diffuse through the hydrophobic core of membrane, from a region of
higher concentration to a region of lower concentration;
2. Osmosis involves movement of water molecules through a partially permeable
membrane. Facilitated diffusion involves movement of polar (or charged) molecules or
ions across the membrane via channel or carrier proteins;

N13P2Q8

(a) Explain the significance to the alpha and beta cells of their blood supply. [3]
1. Alpha and beta cells produce and secrete hormones glucagon and insulin respectively;
2. A good blood supply ensures that the hormones produced by these cells can be secreted
directly into the bloodstream;
3. The hormones will also be able to travel quickly to their target organs where they will
perform their function; [3]

(b) With reference to Fig. 8.2, outline the advantages of such a cell signalling pathway. [3]
1. Facilitate signal amplification
- only a small number of glucagon molecules needed to solicit a large response from
cell
2. Provides multiple checkpoints for regulation
- Several steps in the signalling pathway can be regulated and controlled. E.g.
activation/ inactivation of G protein, activation of adenyl cyclase and amount of cAMP
produced. This will eventually regulate the cellular response of the pathway.
3. Multiple responses to 1 glucagon molecule (ligand)
- 1 glucagon molecule will result in the production of several secondary messengers,
e.g. cAMP.
- cAMP can then trigger multiple signal transduction pathways to elicit different
responses;
4. Ensure specificity
- glucagon has a specific conformation that is complementary to the ligand binding site
of GPCR.
- binding of a specific ligand (glucagon) to a specific receptor (GPCR) will elicit specific
reaction via specific pathway in each cell type;
Raffles Institution Nov 2013 (H2 Biology) Paper 2 (for 9744 syllabus) 2017

(c) Describe how cAMP also increases blood glucose concentration. [3]
1. cAMP is the secondary messenger in the glucagon signalling pathway;
2. Amplification of the signal transduction pathway begins with adenyl cyclase converting
many ATP to cAMP.
3. Numerous cAMPs which are small, non-protein water-soluble molecules are able to
diffuse quickly throughout the cytosol to
4. Activate many other relay molecules/protein kinase A thus leading to the activation of
many enzymes involved in break down glycogen to high glucose production in the liver
cells;
5. The glucose molecules are then released into the bloodstream causing the blood glucose
concentration to increase;

(d) State how the cell signalling pathway for insulin differs from that for glucagon as shown in
Fig. 8.2. [1]
The cell signalling pathway for insulin makes use of a receptor tyrosine kinase (RTK) system
while that for glucagon makes use of a G-protein coupled receptor (GPCR) system.

N13P2Q9
(a) Describe the binomial nomenclature of a species and the basis of hierarchical classification
of species into taxonomic groups. [7]
1. In binomial nomenclature, each species is assigned a 2-part latin name with the genus
followed by the species;
2. e.g. Homo sapiens is the binomial name of humans with Homo being the genus while
sapiens is the species. The genus is capitalized while the species is not. Both names
are underlined separately or italicized;
3. Linnaeus came up with this precise naming system because it avoids ambiguity that
may arise if common names are used instead. e.g. there are many “cats” and there
must be a better way to distinguish the different cats;
4. Being systematic, a newly discovered species can be easily categorized and named;
5. Closely related organisms are grouped together in the same taxon, a group of related
organisms at a particular level;
6. Traditional Linnean classification determines relatedness based on observable
morphological similarities but increasingly, evolutionary relationships are taken into
account as well;
7. The organisms are grouped into a hierarchy of increasingly inclusive taxons/categories;
8. Related species are grouped in the same genus, related genera in the same family and
so on;
9. The hierarchical taxons are species, genus, family, order, class, phylum, kingdom,
domain with the species being the most exclusive group and the domain being the most
inclusive group;

(b) Explain the biological concept of the species. [7]

1. The biological concept of a species defines a species as is group of organisms capable
of interbreeding and producing fertile, viable offspring;
2. A species is thus reproductively isolated from other populations and no longer share a
common gene pool with them;
3. The basis of this concept is that two populations have accumulated sufficient genetic
differences in both allele frequencies and unique mutations that they develop both
prezygotic and postzygotic mechanisms that prevent mating and/or fertilization;
4. This definition cannot be applied to asexually reproducing organisms;
5. This concept cannot be applied to extinct species whose breeding behavior cannot be
observed;
6. For ring species, like the salamanders in California, the adjacent subspecies can
interbreed but the non-adjacent subspecies can’t. Where is the line drawn?;
7. There are some organisms that may physically and physiologically be capable of mating
but, for various reasons, do not normally do so in the wild. e.g. different mating call. It is
disputed whether they are indeed a different species;
Raffles Institution Nov 2013 (H2 Biology) Paper 2 (for 9744 syllabus) 2017

(c) Outline the advantages of molecular methods in classifying organisms. [6]

1. They can be used to compare and categorise species so morphologically
indistinguishable due to convergent evolution or are very closely related;
2. On the other hand, remotely related organisms such as bacteria, humans and sunflower
can also be compared because they share some proteins such as cytochrome c and all
known life is based on nucleic acids. This forms a basis for comparison.;
3. They are objective. Molecular character states are unambiguous as A, C, G and T are
easily recognisable and cannot be confused with another whereas some morphological
characters, such as those based on the shape of a structure, can be less easy to
distinguish because of overlaps between different character states;
4. They are quantitative. Molecular data are easily converted to numerical form and hence
are amenable to mathematical and statistical analysis and hence computation. The
degree of relatedness can be inferred and quantified by calculating the nucleotide
differences between species;
5. Some molecular differences may not be reflected as a difference in the morphological
character and hence may not be picked up by morphological analysis. e.g. a nucleotide
difference in the third base of a codon, or intron.
6. While small genetic differences, may result in major phenotypic differences. In such
instances, vast differences in morphology can exaggerate the evolutionary distance
between two species but not in the case of molecular methods. e.g. in snakes, the loss
of forelimbs and elongation of the body, both radical changes in body form, are due to
mutations in several Hox genes that affect the expression of body patterns and limb
formation.
7. Offers a large set of characters to be studied relatively quickly. Each nucleotide position
can be considered a character with 4 character states, A, C, G and T, and organisms
have millions - billions of nucleotide positions. There are limited morphological
characters that can be studied, e.g number of segments, number of legs, shape of
thorax etc;
8. Amino acid sequences for many proteins and nucleotide sequences for a rapidly
increasing number of genomes can be accessed from electronic databases and used for
comparative study and classification. No physical specimen needs to be preserved;
9. Both living and dead tissue may be used so long as the DNA or protein remains intact
and you do not even need an entire specimen. This increases the accessibility of
studying elusive or rare species;

Teacher’s comments:
Students should not simply lift everything from the notes because this is specific to
classification and not about establishing phylogenetic relationships.

N13P2Q10
(a) Describe how infection by HIV causes disease. [7]
1. The HIV attaches to specific target CD4 cells such as T helper lymphocytes/cells* and
macrophages via its gp120 glycoproteins;
2. Upon entering the cell via fusion, reverse transcriptase* will convert the ssRNA
genome into ds DNA and with the help of integrase*, integrates the viral DNA into the
host genome. Host is now at the latency stage and no symptoms are visible;
3. Integration is random and this may result in the activation of a protooncogene or
inactivation of a tumour suppressor gene which may result in cancer (e.g. Karposi
sarcoma, a rare skin cancer more prevalent in AIDs victims);
4. During the activation of HIV, synthesis of viral glycoproteins gp 120 and 41 occurs and
these will become incorporated into the host cell membrane;
5. allowing the immune system to recognize such cells as infected and become a target of
antibodies causing the host cells to lyse;
6. Budding off causes the progressive loss of host plasma membrane and the cells
eventually die;
7. Or the host cell transcription and translation mechanism has been taken over by the
virus producing viral proteins, compromising the vital functions of the cell and the cell
Raffles Institution Nov 2013 (H2 Biology) Paper 2 (for 9744 syllabus) 2017

eventually dies;
8. AVP regarding how infection kills the host cell directly
9. As the target CD4 cells play a central role in the host immune system, destroying these
cells will result in host immune system being compromised/suppressed; (connection
must be made between CD4 cells and its role in immune system)
10. The host cannot fight off infections from bacteria, viruses and fungi effectively and will
suffer from secondary/opportunistic infections* and may even die from them; (effect
of being immune compromised)
Teacher’s comments:
Students should not focus on the viral life cycle and instead focus on how infection causes
disease. This requires students to draw on their knowledge of retroviruses from the topic of
gene therapy (pt 3) and make inferences within the topic of viruses to come up with pt
5.Students cannot always expect straightforward “describe” questions.

(b) Explain why viruses may be regarded as non-living organisms. [7]

1. Viruses are obligate parasites* which requires a living host to support many of their
functions;
2. They lack the ability to reproduce on their own independently.
3. They do not have a cellular organization such as a cell membrane enclosing cytoplasm
with all the various organelles;
4. They do not show metabolic activity outside the host cell. They do not produce their own
source of energy but instead acquires it from the host cell and this energy is used to
maintain metabolic processes while still inside the host cell;
5. They require host cells to make products such as their coat proteins and their nucleic
acids which they can’t on their own;
6. They do not grow and undergo developmental changes in its life cycle. Once a virus
particle is formed and is released, it is largely inert until a new infection cycle begins.
7. They do not respond to stimuli when outside the host cell. Organisms have specialized
receptors that detect environmental stimuli to allow their cells to adjust metabolism in
response. Viruses do not.

(c) Outline the structure and function of viral nucleic acid. [6]
1. Viral nucleic acid is very diverse. A virus can have either RNA or DNA but never both;
(type)
2. and it can be a double or single stranded, circular or linear, single or
multiple/segmented; (organization)
3. The viral nucleic acid encodes the genetic information that when transcribed and
translated will result in the formation of;
4. Important viral components such as capsid proteins, glycoproteins and viral enzymes;
5. These products are important for the assembly of the viral particles;
6. As well as to allow for the replication of the virus;
7. e.g. RNA dependent RNA polymerase for synthesis of mRNA and genome in influenza,
integrase for integration of viral DNA into the host cell genome in HIV, protease to
cleave the polyprotein of HIV into functional proteins and enzymes;
Raffles Institution Nov 2013 (H2 Biology) Paper 3 (for 9744 syllabus)

Nov 2013 H2 Bio Paper 3

.
N13P3Q1
(a)
State two other ways in which RFLP can be used as a biological tool. [2]
RFLP can be used in the analysis of
1. detection of disease e.g. sickle cell anaemia;
2. DNA fingerprinting* in forensics / paternity testing

(b) (i) Complete Table 1.1 to show the number of restriction sites and the size of the fragments
resulting from digestion of each 15kb DNA molecule, for the three samples. [2]
Table 1.1
BamHI EcoRI BamHI and EcoRI

Number of restriction sites 1 2 3

Size of fragments / kb 4,11 2,3,10 1,2,3,9

(ii) Using the information from Fig. 1.1 and Table 1.1, it is possible to map the 15kb length of DNA
for BamHI and EcoRI.

The restriction map for BamHI is shown in Fig. 1.2.

Complete the restriction map in Fig. 1.3 for both BamHI and EcoRI, by adding the positions of
the EcoRI restriction sites. Indicate the size, in kb, of each fragment. [2]

Teacher’s comments:
The 2 answers below cannot get the 10kb fragment, hence incorrect.

(c) Outline why gel electrophoresis separates DNA fragments. [4]

1. Negatively-charged DNA*;
2. migrates towards the positive electrode/anode when subjected to an electric field / current;
3. Fragments migrate through agarose gel matrix, made up of a meshwork of polysaccharides;
which impedes movement of longer fragments more than shorter fragments;
4. Longer fragments migrate slower compared to shorter fragments;
 Raffles Institution Nov 2013 (H2 Biology) Paper 3 (for 9744 syllabus)

(d) Outline the process of DNA hybridization that allows the RFLP pattern for a particular gene to be
visualized [5]
To detect the RFLP pattern of a gene, after the DNA from is digested with restriction enzyme and the digested
DNA separate using gel electrophoresis

1. ds DNA is denatured / made single-stranded and by alkaline / NaOH solution and transferred
to a nitrocellulose membrane; exactly the same position as they were in the gel
2. The nitrocellulose membrane incubated with a radioactive single stranded DNA probe*,
that is complementary in sequence* to part of the target sequence / gene.
3. DNA fragments containing this part of the target sequence will hybridise to the probe by
complementary base-pairing*;
4. After hybridisation, membrane is washed to remove any unhybridised probes.
5. Using Autoradiography/X-ray film* over the membrane, the banding pattern can be
visualised. (The radioactivity of the bound probes exposes the film to form an image
corresponding to the bands that have base-paired to the probe.)

N13P3Q2
2
(a) (i) Cytosine is a pyrimidine. Name the other pyrimidine. [1]
Thymine*

(ii) Suggest how methylation of cytosine nucleotides prevents the DNA of a prokaryote from being
cut by its own restriction enzymes. [2]
1. Methylation of cytosine in prokaryotic DNA changes the conformation of the DNA at the
restriction site* such that it is no longer complementary in shape and charge* to the
restriction enzyme’s active site*.
2. Hence, the restriction enzyme will be unable to recognise and cleave prokaryotes’ own
DNA.

(b) (OUT OF SYLLABUS)

(c) Another restriction enzyme, BspLI, has the same restriction site

Where N can be any nucleotide.

Using Table 2.1, a scientist predicted that BspLI would cut at more than 20 sites in the standard
DNA.

Suggest why the scientist made this prediction. [2]

1. There are 4 possible nucleotides (adenosine triphosphate/ Guanosine triphosphate /
cytosine triphosphate / thymine triphosphate),
2. Since BspLl restriction site is less specific than of Sfol, there will be more than 20 cut sites;

N13P3Q3

(a) (i) Explain why the gene has at least 200 000 base pairs, but the protein only has 1480 amino
acids. [3]
1. Besides the 4 440 bases which make up exons, code for the 1480 amino acids;
2. there are non-coding DNA such as introns;
3. e.g. promoter, stop codon, UTR (reject control elements such as enhancers/ silencers);

(ii) (OUT OF SYLLABUS)

Raffles Institution Nov 2013 (H2 Biology) Paper 3 (for 9744 syllabus)

(iii) Suggest, with reasons, why other mutations of the CFTR gene vary in the extent to which they
cause symptoms of cystic fibrosis. [3]
1. Single base substitutions which lead to coding of another amino acid with similar R-group
properties will cause little effect on symptoms of cystic fibrosis as it can still fold to similar
conformation as original protein;
2. Mutations such as single base insertion or single base deletion within exons
3. can lead to frameshift mutation leading to an incorrect sequence of amino acid formed with
different R-groups which cannot fold to form a functional protein leading to serious
complications.
4. Base substitutions which lead to a premature stop codon can lead to a truncated protein
which is non-functional will lead to have severe symptoms of cystic fibrosis.

(b) (OUT OF SYLLABUS)

N13P3Q4
Planning Question
1. Aim : To investigate the lowest concentration of copper sulfate solution that has an effect on the
leakage of pigment from beetroot cells.

2. Theory
(Main theory) [Max 1m]
[T1] Betacyanin is found in beetroot cells is prevented from leaking out of cells by membranes
(vacuole & plasma membrane) which are made up of phospholipid bilayer embedded with
proteins
[T2] The betacyanin are too large to pass through the transient pores in the phospholipid bilayer
/The betacyanin are charged/polar and are unable to pass through the non-polar hydrophobic
hydrocarbon core of the phospholipid bilayer.
[T3] Copper ions in the copper sulphate causes denaturation of the membrane proteins embedded
in the phospholipid bilayer. The loss of 3D conformation of the membrane proteins increases
the permeability of the membrane.
Accept when make reference to copper ions disrupting ionic interactions in tertiary structures.

(Measurable quantity)[ 1]
[T4] the degree of permeability of the membrane will be reflected by the amount of betacyanin
pigment leaking out of cells. The permeability is determined by measuring the time taken for
the colour of the bathing solution to match the colour standard.

(Predicted trend) [1]

[T5] Increasing concentration of copper sulfate increases the membrane permeability which results
in a shorter time taken for the colour of the bathing solution to match the colour standard.
[T6] Calculate rate of leaking and extrapolating lowest concentration that has an effect from the
graph

3. Procedure (PAN CR)

a) [P] Pilot test*
Conduct a pilot experiment to determine suitability of apparatus, suitability of range of
independent variable (e.g. concentration of copper sulfate solutions), optimum conditions,
amount of materials used (number of pieces of beetroot cylinders, volume of distilled water) [1]
Raffles Institution Nov 2013 (H2 Biology) Paper 3 (for 9744 syllabus)

b) Annotated diagram

Test tube with 1 beet root

strip (1cm diameter x 5cm
length) and 10 cm3 of
distilled water
Thermometer to monitor
temperature of water bath
and add boiling or tap water
to maintain temperature

500 cm3 beaker

for water bath at
30 C

c) Variables
[DV] Dependent variable AND description of how it is measured : Time taken/s for
solution to match the colour standard [2]

Comment: Dependent variable Rate of leakage of pigment/ s-1 How is this measured 1/time
taken
[IV] Independent variable (at least 5 concentrations at equal interval) AND description of
how it is controlled : 0.2%, 0.4%, 0.6%, 0.8% and 1.0% copper sulfate solution [2]
[CV] Controlled variable and description of how it is controlled: [Max 2m]
Temperature
Type/source/ species of beetroot used
Length /size of beetrootstrip used
Volume of copper sulfate solution used
Any valid point

d) Numbered steps
1. Using a cork borer* (diameter 1cm), cut out cylinders of beetroot*. Using a ruler and scapel,
cut the cylinders of beetroot to a length of 5cm each. (dimension of the beetroot should be
sensible, 10cm x 10cm x 10cm is really too BIG!) Prepare at least 55 beetroot cylinder. (number
of beetroot cylinder require should be sufficient for setting up both the repeats and replicates)

2. Rinse the beetroot cylinders in running water until the water is colourless.

3. To prepare colour standard, place 1 beetroot cylinder in 10cm3 of distilled water in a test tube
and incubate in a water bath of 80oC. After 5 minutes, remove the beetroot cylinder from the test
tube using a pair of forceps and discard it. The red solution left in the test tube will be the colour
standard. [1]

4. Prepare a 30oC water bath by mixing hot and cold water in a 500cm3 beaker. Use a thermometer
to measure the temperature and adjust the temperature to 30oC by mixing hot and cold water.

5. Prepare 10 cm3 0.2%, 0.4%, 0.6%, 0.8% and 1.0% Copper sulfate solution by mixing appropriate
volumes of 1.0% copper sulfate solution and distilled water in a test-tube as shown in the table
below. Use a 10cm3 syringe to measure the volume stated in the table.
Raffles Institution Nov 2013 (H2 Biology) Paper 3 (for 9744 syllabus)

Concentration of Volume of 1% Volume of distilled Total volume

copper sulfate copper sulfate water used /cm3 /cm3
solutions /% solutions used /cm3
0.2 2 8 10
0.4 4 6 10
0.6 6 4 10
0.8 8 2 10
1.0 10 0 10

6. Place the test tubes contain various concentrations of copper sulfate solutions in the 30oC water
bath. [E] Allow 2 min of equilibration time for the temperature in the test tube to reach 30ºC. [1]

Comment: Many students equilibrate by putting the beetroot in the copper sulfate solution. This
is not acceptable. By doing so, the reaction would have started and the proteins in the beetroot
cell membrane are likely to be disrupted within the equilibration time.

7. Place 1 beetroot cylinder into the test-tube containing 0.2 % copper sulfate solution. Start the
stop watch.

8. Stop the stop-watch and record the time taken for the colour of the copper sulfate solution to
matches the colour standard. Use the white cardboard as a background when comparing the
colour of the test-tube and colour standard.

9. [R] Repeat step 5 to 8 using 2 more test tubes of copper sulfate solution with beetroot cylinder to
serve as replicates, to check for anomalous results.[1]

10. [RR] Repeat the entire experiment twice to check for reproducibility of results. [1]

e) [C] Control [1]

Keeping all other variable such as volume and temperature constant, place 1 beetroot cylinder in
10cm3 of distilled water and measure the time taken for the colour of the distilled water to match
the colour standard. This is to show that any change in the permeability is due to the copper
sulfate solution.

Comment: Some students mentioned using glass beads to substitute for the beetroot to show
that red pigments come from beetroot cells. This is not acceptable for this experiment since the
aim of this experiment is to investigate the effects of metal ions on membrane permeability.
Some students wrongly used boiled and cooled beetroots in their control tube. Boiling disrupts
membrane integrity and pigments will leak from the cells.
Raffles Institution Nov 2013 (H2 Biology) Paper 3 (for 9744 syllabus)

4. Data Recording and Processing

Record and process data as follows:
[T] Table showing time taken for the colour of solution to match colour standard [1]
Concentration Time taken for colour of solution to match Rate of
of copper colour standard /s leakage of
sulfate Replicate Replicate Replicate Average pigments /s-1
solutions /% 1 2 3
0.2
0.4
0.6
0.8
1.0
[G] Graph showing the effects of concentration of copper sulfate solution on the time taken for
the colour of solution to match colour standard [1]

Rate /s-1

[1] To obtain the lowest concentration of copper sulfate solution that has an effect on the leakage of
pigment from beetroot cells, extrapolate the graph to obtain the concentration when graph cuts x-axis.

5. [RP] Risks & Precautions [1]

a. When using scalpel, cut the beetroot away from the hand to avoid getting cut.
b. When using the cork borer, use the sharp end on the beetroot to avoid accidentally cut
your hand.
c. Avoid touching the hot Bunsen burner. Only handle it after it has cooled to prevent burns.
d. Use insulating gloves when handling the hot water to prevent scalding.
Raffles Institution Nov 2013 (H2 Biology) Paper 3 (for 9744 syllabus)

N13P3Q5

5 (a) Describe features of zygotic stem cells and embryonic stem cells that distinguish them from
each other. [5]

Point of comparison Zygotic stem cells Embryonic stem cells

Differentiation potential 1a.They are totipotent*; 1b. They are pluripotent*;
Characteristics
2a. They have the ability to 2b. They have the ability to
differentiate* into all of the differentiate* into all of the cell
cell types that make up an types that make up an organism
organism including the extra- except the extraembryonic
embryonic tissue* such as tissue* such as the placenta*;
the placenta*, which
nourishes the embryo;

3a. Hence zygotic stem cells 3b.Embryonic stem cells alone

are able to form the entire cannot form the entire organism
organism; as extraembryonic tissues such as
the placenta is required for foetal
nourishment and development.

Examples and sources 4a. They are derived from a 4b. They are derived from cells of
fertilised egg which forms the the inner cell mass of blastocyst*
zygote. Cells that are at about 4 to 5 days post
produced within the first 3 fertilization;
division (8 cell stage*) after
the egg is fertilized;

(b) Describe the features of blood stem cells and explain their normal functions. [8]

1. Blood stem cells are adult stems cells can be found in the bone marrow* and the
umbilical cord;
2. They undergo self-renewal * by mitotic division to ensure a constant pool of blood
stem cells;
3. As the life span of blood cells are short, blood stem cells replace those lost through
normal cell death.
4. Blood stem cells are multipotent*;
5. They have the ability to differentiate* into several related cell types but is restricted
to blood cells only;
6. They can divide asymmetrically after stimulation by molecular signals;
7. This produces stem cells for the maintenance of the stem cell pool and progenitor cells
to increase or renew the population of specialized blood cells.
8. B and T lymphocytes which are derived from the lymphoid progenitor cell;
9. Red blood cells and platelet producer cells which are derived from the myeloid
progenitor cell;

(c) (OUT OF SYYLABUS )

 Raffles Institution Nov 2014 (H2 Biology) Paper 2 (for 9744 syllabus)

Nov 2014 H2 Bio Paper 2

N14P2Q1
(a) With reference to Fig 1.1, describe the curve between points A and B [2].
1. As temperature increases from 350C (pt A) to 500C, the rate of reaction of
enzyme increases gradually from 0 to 8a.u.
2. However, from 500C to 650C (pt B), there is a steeper increase in rate of reaction
of enzyme from 8a.u. to 65 a.u. This increase is about 7 times that of the
increase in rate from 350C to 500C .

(b) Explain why the reaction rate changes from:

(i) A to B [3]
1. As temperature increase from 350C (at A) to 650C (at B), there is increase in
kinetic energy of the enzyme and substrate molecule.
2. which increases frequency of effective collisions* between substrate and
enzyme thus rate of formation of enzyme-substrate complex* increases ;
3. increasing temperature also increases the number of molecules having sufficient
energy to overcome the activation energy* barrier to form the products of
reaction;
(ii) C to D [3]
1. At 750C (pt C), the optimum temperature* of Taq polymerase, there is greatest
number of molecular collisions and hence max rate of reaction
2. As temperature increase further to 850C (pt D), which is beyond optimum
temperature, thermal agitation / vibration of enzyme molecule breaks hydrogen,
ionic bonds and other weak interactions that stabilizes the 3D conformation
resulting in denaturation*
3. The enzyme active site* no longer complementary in shape* and charge to the
substrate and the rate of reaction decreases steeply from maximum rate of 95
a.u. to almost 0a.u.

(c) Suggest how the structural features of Taq polymerase make it thermostable [2]

1. More Cysteine* residue may be present in the Taq polymerase allowing the formation
of more disulphide linkages* with another cysteine residue to maintain the 3D
conformation
2. Disulphide linkages are strong covalent bonds difficult to be broken by high
temperature
3. Hence the 3D conformation of active side maintained \

N14P2Q2

(a) Give the full name of the bond holding the two glucose molecules together in the way shown.
[2]
1. -1,4-glycosidic bond

(b) Describe how this bond may be broken to release two molecules of glucose. [2]

1. Addition of water in hydrolysis* reaction

2. Enzymatic action of maltase
3. Acid hydrolysis by heating it with hydrochloric acid
(either 2. or 3.)
 Raffles Institution Nov 2014 (H2 Biology) Paper 2 (for 9744 syllabus)

(c) (i) Describe the arrangement of phospholipids in cell membranes. [2]

1. 2 layers of phospholipid molecules in a bilayer

2. With the hydrocarbon tails facing the inside of the membrane and the phosphate heads
facing the outside next to aqueous medium

(ii) Explain how the structure of phospholipids is related to this arrangement in cell
membranes. [3]

1. Phosphate heads are charged and hydrophilic and will form hydrogen with water
2. Hydrocarbon tails are non-polar and therefore hydrophobic and arranged away from
aqueous medium
3. And forms hydrophobic interactions with the hydrocarbon tails of other phospholipid
molecules to form a hydrophobic core in the bilayer structure

[Total : 9]
N14P2Q3

(a) Describe how bacterial chromosomes differ from eukaryotic chromosomes in terms of structure and
organization. [4]
Feature Prokaryotic genome Eukaryotic genome
5 7 7 11
Size 10 -10 base pairs 10 -10 base pairs
Appearance Generally a single, circular Multiple, linear molecules
molecule
Molecule Double Helix DNA
Association with Yes – relatively less Yes – large amounts
proteins e.g. histone-like proteins e.g. histones, scaffold proteins
Level of DNA Relatively low: High:
packing/coiling DNA double helix
DNA double helix
some looping around histone-like
A
proteins Histones
Nucleosome
(“bead”)

Circular chromosomal DNA Linker DNA

(“string”)

Nucleosomes (10-nm fiber) - Euchromatin

B
Formation of looped domains

Looped domains
Nucleosome
30-nm fiber - Heterochromatin
C
Protein scaffold

Supercoiling

D Looped domains (300-nm fiber)

(A) Unfolded chromosome from E. (A) DNA double helix is

coli has a diameter of 430µm. associated Metaphase chromosome
(B) DNA is folded into chromosomal with proteins called
looped domains by protein- histones.
DNA associations. Six domains DNA molecules are
are shown, but actual number is negatively- charged, histone
Raffles Institution Nov 2014 (H2 Biology) Paper 2 (for 9744 syllabus) 2017

about 50. are positively-charged.

(C) Supercoiling cause further DNA thus is held around
compaction, such that it histones by electrostatic
fills an area of about 1 µm. interactions.
Most of DNA is wound
around octamers of 8
histone proteins to
form nucleosomes, the 10
nm fibre. Remainder of DNA,
called linker, joins adjacent
nucleosomes.

(B) The 10-nm fibre coils around

itself to form a 30 nm
chromatin fiber (or solenoid).
(C) The 30-nm fibre forms loops
called looped domains (a
300-nm fibre) when
associated with scaffold
proteins.
(D) Supercoiling present. The
loops further coil and fold to
produce characteristic
metaphase
chromosome.
Location Nucleoid region, not membrane- Nucleus (surrounded by nuclear
bound envelope)
Extrachromosomal Present – plasmids (much smaller No plasmids (however
DNA rings of DNA compared with bacteria mitochondria and chloroplast
chromosome) have their own DNA)
Number of genes 4,500 25,000
Non-coding regions Not common – typically less than Common – about 98%
(between and within 15%
genes)
i. introns None present Many present
(rare; only in some genes)
ii. promoters Present
iii. enhancers/ Rarely present Present
silencers
iv. repeated Few Many
sequences
v. operons Many Few known ones (e.g. in
nematodes)
Origin of replication One Many
(per chromosome)
*Genome refers to a complete set of genetic material in a particular cellular component.
 Raffles Institution Nov 2014 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Describe what occurs from the end of stage 3 up to stage 4, as shown in Fig. 3.1. [4]
(i)
1. After contacting a recipient cell, the sex pilus retracts, pulling the two cells together.

2. the F+ cell then forms a temporary cytoplasmic mating bridge* with the F- cell and
transfers its F plasmid DNA to it.

3. one strand of the double-stranded F plasmid is nicked by a nuclease.

4. The free 3’ end of the nick is extended by DNA polymerase for the synthesis of a new
complementary strand using the intact strand as the template

5. The newly synthesized strand displaces the nicked strand which is transferred
concurrently, via the 5’ end, across the mating bridge into the recipient cell.

6. Upon completion of a unit length of the plasmid DNA (after 1 round), another nick
occurs to release the original strand and end the replication of the newly synthesized
strand;

(ii) Explain the role of F plasmid. [3]

1. F factor on F plasmid, codes for proteins necessary for the formation of sex pili and
subsequent cytoplasmic mating bridge,
2. allowing for conjugation to occur between bacteria.
3. This allows for bacterial genes to be transferred between bacteria and hence increases
genetic variation between bacteria.
[Total : 11]

N14P2Q4
(a) Suggest the reason for the peak in the rate of DNA synthesis shown in Fig. 4.1. [2]
1. Gene coding for ribosomal RNA was amplified;
2. To allow for an increase/high rate of ribosomal RNA synthesis during egg cell
growth;

(b) Describe and explain the pattern of transcription visible on the part of the DNA coding for
ribosomal RNA, labelled X in Fig. 4.2. [2]

1. Multiple ribosomal RNA transcripts are produced concurrently from transcription

of a single gene,
2. and length of the ribosomal RNAs increase progressively from left to right of
region X ;
3. This is because the direction of transcription takes place in a single
direction/ribosomal RNA is produced in 5’ to 3’ direction;

c) Explain why such large amounts of ribosomal RNA are required in frog egg cells. [2]

1. Large amounts of ribosomal RNA are required for synthesis of large numbers of
ribosomes in the frog egg cells;
2. This is required for high rate of protein synthesis following fertilisation of the egg
cells;
 Raffles Institution Nov 2014 (H2 Biology) Paper 2 (for 9744 syllabus) 2017

d) Suggest possible roles for the non-transcribed DNA that is found between the ribosomal
genes. [2]
1. the non-transcribed DNA contain promoter*;
2. which binds RNA polymerase* and general transcription factors* to form the
transcription initiation complex* for transcription to take place;
OR
3. the non-transcribed DNA may contain enhancer*;
4. which binds activators* to promote the assembly of general transcription
factors* and RNA polymerase* at the promoter* of the ribosomal gene to form
the transcription initiation complex*;
OR
5. the non-transcribed DNA may contain silencer*;
6. which binds repressors* to prevent the assembly of general transcription
factors* and RNA polymerase* at the promoter* of the ribosomal gene to form
the transcription initiation complex*;
e) Explain how operons allow for rapid response by bacteria to environmental change. [2]
1. multiple genes coding for products involved in the same biochemical pathway are
under the control of a single promoter*;
2. these genes are therefore turned “on” or turned “off together, allowing the
bacteria to respond rapidly to changes in the environment;
[Total : 10]
N14P2Q5
5(a) Draw a genetic diagram to explain both crosses. [5]
Use the following symbols to represent the different alleles involved:
R -red flowers r -white flowers A -axial flowers a -terminal flowers
Parental phenotype Red, axial flowers X White, terminal flowers
Parental genotype RRAA X rraa

Gametes RA ra

F1 phenotype RrAa [1]

F1 phenotype All red, axial flowers [1]
F1 selfing RrAa x RrAa

Gametes
RA Ra rA ra

RA RRAA RRAa RrAA RrAa

Red, axial Red, axial Red, axial Red, axial

RRAa RRaa RrAa Rraa

Ra Red, axial Red, terminal Red, axial Red, terminal

rA RrAA RrAa rrAA rrAa

Red, axial Red, axial white, axial white, axial

ra RrAa Rraa rrAa rraa

Red, axial Red, terminal white, axial White, terminal

Offspring genotype R_A_ R_aa rrA_ rraa

Offspring phenotype Red, Red, White, White,
axial flowers terminal flowers axial flowers terminal flowers
Phenotypic ratio 9.5 3.1 2.7 1
9 3 3 1
[1] correct offspring genotypes in Punnett square
[1] relate genotype to phenotype
[1] correct phenotypic ratio of 9:3:3:1
 Raffles Institution Nov 2014 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Explain how different characteristics can be inherited independently in dihybrid inheritance. [2]
1. 9: 3: 3: 1 offspring/F2 phenotypic ratio indicates the alleles of each character are found on
different chromosomes and inherited independently of one another;
2. The alleles of different genes assort independently* of each other because homologous
pairs of chromosome align randomly on either side of the metaphase plate/ because
alignment of each homologous pair is independent of other homologous pairs/ because two
alleles for flower colour segregate independently of the two alleles for flower position.

(c) Explain why heterozygous plants for this gene, Tt, have the same phenotype as homozygous
dominant plants, TT. [3]

1. Allele T codes for a functional enzyme/is the functional copy of the gene involved in
synthesis of gibberellic acid.
2. Allele t codes for the non-functional enzyme/ is the non-functional copy of the gene.
3. In a heterozygote, allele T masks the presence of allele t.
4. Presence of 1 T allele (in homozygous dominant or heterozygous individuals) causes T
allele to be expressed, producing functional enzyme that causes plants to be grow tall.
(Allele T is dominant over t)
[Total : 10]
N14P2Q6
(a) Name the structures labelled A, B, C, and D on Fig. 6.1. [4]
A Intergranal lamella
B Chloroplast envelope (R: inner membrane or outer membrane)
C Stroma
D Granum
b(i) Suggest two advantages of these large protein complexes being held in the membranes of
structures A and D in Fig. 6.1. [2]
1. Maintains the sequential arrangement of the photosystems* and electron carriers of
electron transport chain* for the flow of electrons;

2. Maintains proton gradient for ATP synthesis since the hydrophobic core* of the
membrane is impermeable to protons and this is essential for chemiosmosis
OR
The enclosed thylakoid space allows the build up of protons here by proton pumps found in
the electron transport chain, enabling the establishment of a proton gradient across the
thylakoid membrane for chemiosmosis.

3. Allows ATP synthase to be embedded in the correct orientation, where the active site faces
the stroma / for protons to diffuse from thylakoid space to stroma.

4. Through compartmentalization, it improves the efficiency of the different reactions by

bringing the large molecules involved in photophosphorylation closer together
5. The thylakoid membrane is highly folded, provides a large surface area to embed
Any of the elaboration:
many photosynthetic pigments / chlorophyll molecules in the photosystems for light
absorption*
many electron carriers in the electron transport chain and ATP synthase for ATP
production.

(ii) Outline the role of ATP synthase that is held in these membranes. [2]
1. Diffusion of protons from thylakoid space into the stroma, via ATP synthase
2. Leads to formation of ATP from ADP and inorganic phosphate via chemiosmosis*.
 Raffles Institution Nov 2014 (H2 Biology) Paper 2 (for 9744 syllabus) 2017

(c) Describe the role of NADP in linking the light dependent reactions to the Calvin cycle. [2]
1. NADP is the final electron acceptor* at the end of the non-cyclic light dependent reaction,
which is then reduced to NADPH.
2. Reduced NADP/NADPH is used to drive the reduction of glycerate phosphate (GP) to
glyceraldehyde-3-phosphate (G3P), with the use of ATP in the Calvin cycle.
[Total : 10]

N14P2Q7
(a) Describe events occurring at A and B in Fig. 7.1. [4]
Stage A
1. Haemagglutinin* binds to sialic acid receptors* on host cell membrane
2. Virus then enters host cell by endocytosis* where host cell membrane invaginates and
pinches off, placing virus in an endocytic vesicle.
Stage B
3. Vesicle then fuses with a lysosome, and the resulting drop in pH stimulates fusion of viral
envelope with vesicle membrane
4. Releasing nucleocapsid and eventually viral RNA into cytosol of host cell
(b) Explain why it is necessary for the viral RNA to enter the host cell nucleus. [3]

1. For transcription* where the viral genome RNA* is used as a template* to synthesise the
mRNA* strands catalyses by the viral RNA dependent RNA polymerase*
2. Which can be transported into cytosol as a template* for translation of viral proteins
3. which in turn acts as a template* for the replication of new viral RNA genome
4. Using the host cell’s ribonucleotides (and ATP), which is abundant in nucleus, for its
synthesis

(c) Suggest in outline how new strains of influenza virus may arise. [3]
1 New strains of viruses are formed as a result of mutation of the genome due to the change
in the ribonucleotide sequence known as antigenic drift*
2 As a result of the lack of proof reading ability of RNA-dependent RNA polymerase in
influenza / the fast/high rate of replication of the virus
3 gives rise to changes in the conformational of the glycoproteins
4 New viral strains are also formed when two or more strains of influenza viruses infect a
common/same host cell where
5 Reassortment of the different RNA segments occur resulting in recombination of genetic
material in a virion*, giving rise to antigenic shift*.
[Total : 10]
N14P2Q8
(a) Suggest why different species of picture-wing fly show different banding. [3] 6L
1. (Phenotypic differences/different wing patterns) distinguishes species in picture-wing
flies;
2. Such differences are due to differences in DNA sequences/mutations in several gene
loci;
3. These changes in DNA structure are reflected by the different staining of DNA /
different banding patterns in the chromosomes;
 Raffles Institution Nov 2014 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Suggest why different species of picture-wing fly evolved on different islands. [4] 8L
1. The islands were geographically isolated with the sea separating the islands/allopatric
speciation (with mention of type of barrier);
2. This meant that the gene flow between populations of flies on the islands were
disrupted*;
3. With the different environment with their different selection pressures*, e.g. predation,
habitats, available food sources (name one example);
4. Allele frequencies change because of natural selection and genetic drift*;
5. The subpopulations evolve independently and accumulate different mutations and allele
frequency changes, that over time led to reproductive isolation and formation of different
species;

(c) Explain the differences between classification and phylogeny. [3]

Fig.8.3 was shown to direct you towards reasons related to the branching network of a
phylogram. Any 3
1. Classification groups organisms based on overall/morphological similarities and does
not take into account the evolutionary history of organisms while phylogeny traces
the evolutionary history of organisms based on ancestor-descendent relationships;
2. Classification involves grouping organisms into a kingdom, phylum, class, order,
family, genus and species using a hierarchical classification* system but in
phylogeny, organisms are arranged based on their evolutionary relationship with
each other with each organism assigned a position on a branching tree relative to
other organisms;
3. In classification, the organism is presented as a binomial nomenclature/name but in
phylogeny, the organism is placed on a branch on a phylogenetic tree;
4. Classification does not allow inference of speciation events as it is just a naming
system while phylogeny indicates speciation events as the node of the phylogenetic
tree;
5. Classification does not allow inference of common ancestors while in phylogeny,
descendants of a common ancestor are represented in the same branch;
6. You cannot infer of how closely related 2 species are especially since they are
grouped together in the same hierarchy, e.g. “species” in classification. In phylogeny,
how recently a branch point occurs indicates how closely related 2 species are;
[Total : 10]
N14P2Q9 (OUT OF SYLLABUS)
 Raffles Institution Nov 2014 (H2 Biology) Paper 3 (for 9744 syllabus)

Nov 2014 H2 Bio Paper 3

N14P3Q1
1(a) (i) Name one genetic disease which has been treated with stem cell transplantation. [1]

Bone marrow haematopoietic stem cells transplants from normal healthy bone marrow
donors to leukaemia patients
Or
Neural stem cell transplant for Parkinson’s disease/multiple sclerosis by introducing adult
neural stem cells into damaged tissue.

(ii) Explain why stem cell is suitable for this purpose. [3]

1. Adult stem cell is multipotent that differentiates* into the respective specialized* cell
type, thus restoring function of damaged or diseased tissue.
2. Self-renewing* nature of stem cells ensures that transplanted stem cells constantly
replicate* in the patient to maintain a constant pool of stem cells
3. As the ‘healthy’ stem cell carries the normal and functional allele thus they can
produce normal levels of functional protein and be used to treat genetic diseases

b(i) Using the letter R, label Fig. 1.1 to identify a feature that allows the virus to bind to cells.
[1]
Note: R can be any one of the 2 types of glycoproteins, one arrow is sufficient.

(ii) With reference to your knowledge of retroviruses, explain how expression of an inserted
gene (transgene) is brought about following infection of host cells with the lentiviral
vector. [3]
1. Once inside the host cell, retroviruses create double stranded DNA copies of their
RNA genomes via reverse transcriptase*.
2. Viral genome together with the transgene are integrated randomly into host
chromosomes via integrase.
3. Transgene will undergo transcription and translation by host enzymes to produce
normal, functional protein.

(iii) (OUT OF SYLLABUS)

(c) (OUT OF SYLLABUS)

[Total : 12]
N14P3Q2 (OUT OF SYLLABUS)
 Raffles Institution Nov 2014 (H2 Biology) Paper 3 (for 9744 syllabus)

N14P3Q3
(a) Describe the limitations of PCR. [3]
Feature Limitations
Taq polymerase lacks 3’ to Errors occurring early in the PCR reaction will get
5’ proofreading ability compounded with each replication cycle and all
daughter molecules resulting from this early error will
be exponentially affected.
Synthesis of PCR primers Success of PCR requires knowledge of
depends on sequence sequences flanking target region to be amplified.
information from target If the flanking sequences of a gene of interest are
region unknown, no proper primers can be synthesised to
amplify the target DNA sequence. If primers are
designed incorrectly, no amplification occurs / wrong
DNA fragment(s) may be amplified.
Limit to size of DNA DNA fragments to be amplified are limited to about
fragment to be amplified 3 kb1. Further increase in length of target sequence
decreases efficiency of amplification. This is because
the polymerase tends to ‘fall off’ DNA template
before chain extension is complete.
Exponential amplification of It is possible to contaminate a fresh PCR reaction
contaminant DNA with minute amounts of contaminant DNA due to
poor laboratory skills. Such unwanted DNA
sequences may be amplified to significant amounts,
alongside the target DNA sequences.

(b) (i) Describe the role of the buffer solution in the gel electrophoresis protocol. [2]

1. Buffers contain ions which allows conduction of electric current

2. Thus allowing the negatively charged DNA molecules to move from the negative
electrode to the positive electrode

(b) (ii) Describe the role of the loading or tracking dye in the gel electrophoresis protocol. [3]

1. Contains glycerol which makes the DNA sample denser than buffer so that DNA
sample can sink to the bottom of the well.
2. DNA is invisible, so the dyes colour the DNA sample showing if it has been loaded
correctly into well.
3. 2 coloured dyes act as visual markers to show the progress of migration of DNA
fragments in the gel.
4. One dye typically (moves at speed corresponding to 100bp DNA fragment which)
runs ahead of sample and another (moves at speed of 1100bp DNA fragment which)
runs after sample.
 Raffles Institution Nov 2014 (H2 Biology) Paper 3 (for 9744 syllabus)

(c) (i) Outline the process of genetic fingerprinting using RFLP that could be used to test this
seized ivory. [4]

1. Genomic DNA is extracted from the soft tissue/dried blood and cut with same
restriction enzyme* to obtain different-sized DNA fragments.
2. DNA is separated according to size in gel electrophoresis where negatively-
charged DNA* migrates towards the positive electrode/anode when subjected to an
electric field / current;
3. Meshwork of agarose fibres impedes movement of longer fragments more than
shorter fragments resulting in smallest fragments moving furthest/largest fragments
least far from well
4. ds DNA is denatured / made single-stranded and by alkaline / NaOH solution and
transferred to a nitrocellulose membrane
5. Carry out Southern blotting/nucleic acid hybridisation by incubating membrane with
single strand radioactive probe* which will hybridise with DNA fragment through
complementary base pairing
6. Using autoradiography/X-ray film* over the membrane, the banding pattern can be
visualised.

(ii) Explain how the genetic fingerprints of the seized ivory could be used to confirm that it
originated from elephants in Malawi. [4]

1. Genetic fingerprint is due to different alleles/markers producing different bands in

gel resulting in the unique banding pattern in individuals
2. Different bands arise due to polymorphic nature of DNA in different individuals,
there will be variations in number and location of restriction sites and number of
tandemly repeated nucleotide sequence among individuals.
3. Genetic fingerprint of animals that provided ivory can be compared against
fingerprint of elephants from Malawi to see how closely related they are.
4. If fingerprint pattern is similar to pattern to that of Malawi elephants, then ivory haul
from Malawi.

N14P3Q4

4 Planning
[Total: 12]

Suggested answer scheme:

Part 1: Aim
To investigate effect of temperature and pH on rate of sucrase activity of 2 enzymes P
and Q.
Part 2: Theory
(Main Theory): [T1: 1 mark for any 2 points from 1 to 5]
1. Active site of sucrase has a specific conformation, complementary in shape and
charge to substrate, sucrose.
2. Conformation of active site, hence rate of sucrase activity, is affected by temperature
and pH.
3. Excess [H+] or [OH-] ions disrupt ionic, hydrogen bonds, which determine
tertiary/quarternary structure of enzyme hence its any conformation.
4. Increasing temperature (up to denaturation) increases kinetic energy of molecules,
increasing rate of effective collisions between enzymes and substrate to form
enzyme-substrate complex.
5. At optimum temperature and pH of each enzyme, rate of production of reducing
sugars is maximum.
(Measurable variable):
Raffles Institution Nov 2014 (H2 Biology) Paper 3 (for 9744 syllabus)

6. Mass of brick red precipitate formed when products (glucose, fructose) obtained from
hydrolysis of substrate (sucrose) are tested with Benedict’s solution. [T2: 1 mark]

Dependent variable: rate of sucrase activity (P and Q) as indicated by rate of brick red
precipitate formed

Independent variable: temperature

(to ascertain optimum temperature)

Part 3: Procedure
a. Pilot Test
Conduct a pilot experiment to determine suitable range of independent variables used,
suitability of apparatus, concentration of substrate (sucrose). If substrate is too
concentrated, it may be diluted with distilled water* (we can’t think of where else to use
the distilled water!) [P: 1 mark]
b. Annotated diagram
Set-up simple, diagram probably not needed.
c. Numbered steps in procedure
1. Fill 3 test tubes with 5 cm3 of 5% sucrose* buffered at pH 7. Use a pH probe, digital
meter to measure and monitor pH, and syringes to measure volumes.
2. Fill another 3 test tubes with 1 cm3 of 2% sucrase P*. These serve as replicates to
check that no anomalies are present. [R1: 1 mark for both replicates and repeats,
including how they are carried out and why they are carried out]
Each sample will follow this procedure:
3. Constant variables include:
volume of substrate - kept constant 5 cm3 because extent of enzymatic reaction is
affected by substrate quantity,
duration of heating in boiling water bath - kept constant 3 minutes during
Benedict’s test as excessive heating converts sucrose into fructose and glucose.
[CV1 and CV2: 1 mark each for 2 constant variables, including what
variables, details on how they are kept constant and why it is necessary to
keep variables constant]
4. Place all tubes in a water bath. Keep temperature constant at 30 C. Maintain
temperature using thermostatically-controlled water bath* and monitor
temperature with a thermometer*.
5. Allow substrate (step 1) and enzyme tubes (step 2) to acclimatize separately for 2
minutes to reach set temperature. Add sucrase to each of the sucrose solutions. Use
stop watch* to check starting time. [E: 1 mark for correct acclimatization with
reason]
6. After reaction has proceeded for 5 minutes, add 6 cm3 Benedict’s solution* to each
tube, place in a boiling water bath over Bunsen burner* for 3 minutes.
7. Pre-weigh each filter paper. After the 3 minutes filter the precipitate. Dry precipitate in
a desiccator till a constant weight. Weigh mass of precipitate.
8. Calculate dependent variable which is rate at which sucrase P hydrolyses sucrose to
fructose and glucose:
average mass of brick red precipitate / time.
[DV: 1 mark – show how to obtain dependent variable including calculation,
method of obtaining must be scientifically sound]
9. Repeat steps 1-8 using buffered sucrose solutions at 20oC, 40oC. 50oC, 60oC. [IV1: 1
mark - independent variable, with total of 5 temp values of equal intervals.
Include how the temperature is created and maintained.]
10. Repeat steps 1-9 twice more to check for reproducibility.
[R1: 1 mark for both replicates and repeats, including how they are carried out
and why they are carried out]
d. Control
Keep all variables constant. Set up control experiment at each of the five
Raffles Institution Nov 2014 (H2 Biology) Paper 3 (for 9744 syllabus)

temperatures using boiled and cooled sucrase.

No precipitate formed for Benedict’s test – shows hydrolysis of sucrose to
monosaccharides is an enzyme-catalysed reaction.
[Co: 1 mark for either control, including how it is carried out and reason why it
is performed]

Part 4: Data recording and processing:

Table showing rate of sucrase P activity at pH 7
[T1: 1 mark for any full table including correct units - either for enzyme P or Q
/ either pH or temperature]
Temperature Mass of precipitate formed in 5 min / g Rate of sucrase
/ oC Replicate 1 Replicate 2 Replicate 3 Average P activity
/ g min -1
20
30
40
50
60

11. Repeat Parts 3 and 4 using sucrase Q to replace sucrase P to determine optimum
temperature of sucrase Q.

Independent variable: pH
Once optimum temperatures for sucrase P and Q are obtained, proceed to identify the optimum
pH of sucrase P and Q.
Repeat Parts 3 and 4 – with the following changes:

Part 5: Procedure
c. Numbered steps in procedure
1. Fill 3 test tubes with 5 cm3 of 5% sucrose buffered at pH 7.
4. Place all tubes in a water bath, keep temperature constant at optimum temperature
of 2% sucrase P* determined in Parts 3, 4.
10. Repeat steps 1-9 using buffered sucrose solutions of pH 5, 6, 8 and 9.
[IV2: 1 mark - independent variable, with total of 5 H values of equal intervals.
Include how the pH is maintained.]
d. Control
Keep all variables constant. Set up control experiment at each of the five pH
using boiled and cooled sucrase.
No precipitate formed for Benedict’s test as in Part 3.
[Co: 1 mark for either control, including how it is carried out and reason why it
is performed]

Part 6: Data recording and processing:

Table showing rate of sucrase P activity, at optimum temp.
pH Mass of precipitate formed in 5 min / g Rate of sucrase P
Replicate 1 Replicate 2 Replicate 3 Average activity / g min -1
5
6
7
8
9

11. Repeat Parts 5 and 6 using sucrase Q replacing sucrase P.

[Fa: 1 mark for doing experiment 1 variable at a time]

Raffles Institution Nov 2014 (H2 Biology) Paper 3 (for 9744 syllabus)

[M: 1 mark for using all compulsory reagents and apparatus marked with *]

Graph showing rate of sucrase activity at pH 7, of enzymes P, Q

Rate of sucrase
activity / g min -1

Sucrase P Sucrase Q

Temperature
/ C
20 30 40 50 60

Graph showing rate of sucrase activity, at the respective optimum temp. of sucrases P, Q

Rate of sucrase
activity / g min -1
Sucrase P Sucrase Q

5 6 7 8 9
pH

[G: 1 mark for either graph with trend and labels]

10. Risks and precautions [1 mark]

(i) Use a boiling tube holder to remove boiling tubes from boiling water bath, in order to
prevent scalding.
(ii) Wear gloves when measuring enzymes P, Q (or use of acid/alkali) as they are
irritants.
Raffles Institution Nov 2014 (H2 Biology) Paper 3 (for 9744 syllabus)

N14P3Q5
(a) and (b) (OUT OF SYLLABUS)
(c) Eukaryotic genes cannot be expressed directly in the bacterial plasmids [7]
because of difference between prokaryotes and eukaryotes, including the
presence of introns.

Outline these problems and explain how they are overcome in order to allow
expression of eukaryotic genes in plasmids within E. coli cells.

1. Pre-mRNA produced from transcription of eukaryotic genes have

introns and exons;
2. During splicing* in eukaryotic cells, introns are excised and exons
joined together by spliceosomes* to form mature mRNA*;
3. Unlike eukaryotic cells, prokaryotic cells do not have spliceosomes*
and are unable to carry out splicing to form mature mRNA;
4. In turn, translation of pre-mRNA in prokaryotes results in non-functional
protein produced;
5. To overcome this problem, reverse transcriptase* can be used to
reverse transcribe mature mRNA* from eukaryotes to form
complementary DNA*;

6. RNA polymerase* in prokaryotes are unable to recognize and bind to

eukaryotic promoters* to express eukaryotic genes;
7. This can be overcome by inserting the eukaryotic gene just
downstream of a prokaryotic promoter* in the plasmid;

8. Prokaryotes are unable to carry out post-translational-modification to

form functional eukaryotic proteins;
9. e.g. functional human insulin is formed following cleavage of the C
chain and formation of disulfide bonds between the A and B chains;
10. A and B chains can be purified separately from different bacteria
cultures and formation of disulfide bonds between them chemically
induced;
 Raffles Institution Nov 2015 (H2 Biology) Paper 2 (for 9744 syllabus)

Nov 2015 H2 Bio Paper 2

N15P2Q1
(a) Identify the structures A and B, as shown in Fig.1.1. [6]
For each structure, state two features that can be seen in Fig.1.1.

structure A: mitochondrion (R: mitochondria)

feature 1: double membrane
feature 2: highly folded inner membrane/cristae
structure B: rough endoplasmic reticulum
feature 1: flattened sacs called cisternae studded with ribosomes
feature 2: continuous with outer membrane of nuclear envelope

(b) Describe two functions of the Golgi body.[2]

1. To glycosylate proteins and lipids to form glycoproteins and glycolipids respectively;
2. To modify existing glycoproteins and glycolipids by modifying/cleaving the existing
sugar chains;
3. To sort and package proteins into different vesicles and target the proteins to
different parts of the cell or for secretion;
4. To form lysosomes;
5. To synthesise polysaccharides such as pectin which is transported in vesicles to the
cell membrane;

(c) Suggest two advantages to eukaryotic cells of having membrane bound organelles.[2]
Membranes allows for compartmentalisation which allow
1. unique environments to be formed for highly specialised activities
(e.g. acidic environment in lysosomes for hydrolytic enzymes to work);

2. spatial separation of biochemical processes & thus their sequential operation

within a cell (e.g. protein modification in RER and further protein modification,
sorting and packaging in the GA) ;

3. accumulation of ions to high concentrations (e.g.accumulation of a high

concentration of H+ in the intermembrane space of the mitochondria enable a
proton gradient to be established for chemiosmosis);

4. Membranes act as a surface for chemical reactions to occur in a sequential manner

membranes may have functionally-related proteins grouped together so that
sequential biochemical processes can occur (e.g. the thylakoid membranes of the
chloroplast have electron carriers & ATP synthetase for chemiosmosis to occur);

5. Membranes increase surface area for chemical reactions (e.g. inner mitochondrial
membrane is highly folded to hold more electron transport chains and ATP
synthetase);

(d) Explain the role of glycogen in animal cells.[2]

1. Glycogen is an large energy store found in the liver and muscles;
2. which can be hydrolysed to many glucose molecules that can be used as a
respiratory substrate which oxidized during respiration to produce ATP.
[Total :12]
 Raffles Institution Nov 2015 (H2 Biology) Paper 2 (for 9744 syllabus)

N15P2Q2
(a) Describe how replication of the lagging strand template occurs. [2]
1. The lagging strand is synthesised discontinuously in fragments known as
Okazaki fragments. Each fragment is initiated by an RNA primer before the
addition of DNA nucleotides;
2. A different DNA polymerase then excises the RNA primer and replaces it with
deoxyribonucleotides and DNA ligase seals the nicks by forming phosphodiester
bonds between adjacent nucleotides of the each of the DNA fragments on the
new strand;
Comments: Do note that in Fig. 2.1: DNA polymerase* works only in the 5’ to 3’ direction.
So the DNA polymerases extending new strands in opposite directions with respect to the
replication fork. i.e. The leading strand is being synthesized towards the replication fork while
the lagging strand is being synthesized away from the replication fork due to the anti parallel
nature of the 2 template strands.
(b) State 2 ways in which DNA replication,
(i) differs from transcription,

DNA replication Transcription

1. Product Double-stranded DNA Single-stranded mRNA
2. Enzymes DNA polymerase links RNA polymerase links
nucleoides nucleotides
3. Primer RNA primer is required RNA primer is NOT required to
requirement to initiate DNA initiate DNA replication
replication

(ii) is similar to transcription.

1. Both use DNA as a template to synthesise the complementary strand;
2. Both processes occur in the nucleus;
3. Nucleotides in the nucleic acid that is synthesized are linked by phosphodiester
bonds;
 Raffles Institution Nov 2015 (H2 Biology) Paper 2 (for 9744 syllabus)

(c) The symbols below represent the main components of RNA.

In the space below, draw a short section of mRNA that is made up of two nucleotides, using
these symbols to represent the main components. Add lines to show the positions of any
bonds between the components.
Label and name the components and the covalent bond that links the nucleotides.[3]

Phosphate group Nitrogenous base

Ribose sugar
group

Phosphodiester
bond

1. Phosphate group linked to carbon number 5 of ribose sugar and nitrogenous base linked to
carbon number 1 of ribose sugar;
2. Correct labels for phosphate group, ribose sugar and nitrogenous base;
3. Phosphodiester bond correctly identified and labelled

[Total : 10]

N15P2Q3

(a) (i) Name structures S and T, as shown in Fig. 3.1. [2]

1. S: promoter*;
2. T: operator*;

(ii) Identify a structural gene in Fig. 3.1 and explain what is meant by the term, structural
gene. [1]
1. lacZ / lacy / lacA gene +
Structural gene is any gene that codes for a protein product that has an
enzymatic function in a metabolic pathway;

(iii) Identify a regulatory gene in Fig. 3.1 and explain what is meant by the term, regulatory
gene. [1]
1. lacI gene +
Regulatory gene codes for a protein (e.g. repressor) involved regulating
expression of structural genes;
 Raffles Institution Nov 2015 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Using Fig. 3.1, describe how the presence of lactose induces a bacterium to use lactose as a
respiratory substrate. [3]
1. Lactose is converted to its isomer allolactose which acts as an inducer to bind to
the active repressor protein at its allosteric site;
2. This makes the repressor inactive as it alters the conformation of the DNA-
binding site of the repressor and which then can no longer bind to the operator;
3. RNA polymerase is now free to bind to the promoter and can move downstream
to transcribe* the structural genes to form -galactosidase, permease and
transacetylase for metabolism of lactose;
[Total: 7]

N15P2Q4
Fig. 4.1 shows the development of a metastatic cancer in the colon over a period of ten or
more years. Metastatic is a term used to describe cancer that is spreading from one organ to
another. APC, ras and p53 are tumour suppressor genes. (error: ras is proto-oncogene!!)***
(a) State two environmental causes of cancer. [2]
1. ultraviolet light / radioactivity / ionizing radiations (mention of radiation alone not
sufficient as visible light also emits radiation);
2. Carcinogens such as tar in cigarette smoke, asbestos, benzene, formaldehyde,
ethidium bromide etc. (give named example);

(b) With reference to Fig. 4.1, explain why cancer development is a multi-step process. [3]
1. The development of cancer requires the accumulation of mutations in the genes
in a single cell; [1]
Idea of accumulation of different mutations using Fig. 4.1 (1mark for any two
points 2,3,4)
2. Loss-of- function mutation in 2 copies/alleles of APC tumour suppressor gene
results in dysregulation of cell cycle to have excessive cell division;
3. Gain-in-function mutation in just one copy/allele of ras proto-oncogene to ras
oncogene results in hyperactive/excessive ras protein to form tumour/mass of
cells;
4. Loss-of- function mutation in 2 copies/alleles of p53 tumour suppressor gene
results in further dysregulation of cell cycle to have excessive tumour/mass of
cells;
5. Chromosomal aberrations and other events (such as activation of telomerase,
loss of contact inhibition, angiogenesis) occur to eventually lead to metastasis; [1]

(c) Describe how dysregulation of the checkpoints of cell division may lead to cancer. [2]
1. Ref to dysregulation of any one of the three checkpoints in cell cycle:
M checkpoint dysregulated thus if any chromosomes are not attached to spindle
fibres, the cell continues into metaphase and anaphase to produce genetically
altered cell/mutant cell;
OR
G1 / G2 checkpoint dysregulated thus damaged DNA not repaired and cell
continues into the M phase, accumulating the mutations;
2. Uncontrolled cell division / excessive cell division that leads to tumour/a mass of
cells;
 Raffles Institution Nov 2015 (H2 Biology) Paper 2 (for 9744 syllabus)

(d) Outline the role of tumour suppressor genes in the development of cancer. [3]
1. Loss-of-function mutation of tumor suppressor genes causes no functional
gene products/proteins to form;
As a result,
2. unable to stop cell cycle to allow repair any damaged DNA;
3. unable to activate DNA repair mechanism to repair damaged DNA thus
accumulation of mutations occurs;
4. unable to initiate/promote apoptosis thus cell with potential to cause cancer is not
removed;
[Total: 10]

N15P2Q5

In guinea pigs, the black coat allele B is dominant to the white coat allele b, and the straight
hair allele H is dominant to the wavy hair allele h.
A guinea pig with a black coat and straight hair was crossed with a guinea pig with a white coat
and wavy hair. The resultant offspring all had black coats with straight hair.
All these offspring were then crossed with guinea pigs having white coat and wavy hair in a
series of test crosses.
The following progeny were produced from the test crosses :
Black coat, straight hair 30
Black coat, wavy hair 10
White coat, straight hair 12
White coat, wavy hair 31
Total 83
(a) Using the symbols for the alleles stated above, draw a genetic diagram t show the expected
phenotypic ratios for the offspring of the test crosses if the inheritance is Mendelian.
Parental
Black coat, straight hair X White coat, wavy hair
phenotype

Parental genotype BbHh X bbhh

Gametes BH Bh bH bh bh

BH Bh bH bh
BbHh Bbhh bbHh bbhh
bh Black coat Black coat white coat white coat
straight hair wavy hair straight hair wavy hair

Offspring BbHh Bbhh bbHh bbhh

genotype
Offspring Black coat Black coat white coat white coat
phenotype straight hair wavy hair straight hair wavy hair
Phenotypic ratio 1 1 1 1

1m for correct parental phenotype and genotype ;

1m for correct gametes which are circled ;
1m for correct offspring genotype and phenotype ;
1m for correct phenotypic ratio ;
 Raffles Institution Nov 2015 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Explain why there is a greater number than expected of the parental phenotypes. [3]
1. The genes coding for coat colour and hair texture are linked on the same chromosome;
2. Alleles for black coat and straight hair are linked on the same chromosome;
3. Alleles for white coat and wavy hair are linked on the same chromosome;
4. Greater chance for these alleles to be inherited together, thus resulting in greater
number;
5. Number of recombinants are smaller because crossing over/recombination is a
chance event and the frequency of recombination is dependent on the distance
between the two genes;

(c) Describe how it is possible for progeny with black coats and wavy hair to be produced from
these test crosses. [3]

1. During prophase I of meiosis I, crossing over* occurs between non-sister

chromatids of homologous chromosome*;
2. At the chiasma*, portion of chromatid containing allele B break and rejoin to portion of
chromatid containing allele h;
3. Resulting in new linkage group being formed where the allele B that codes for black
coat and the allele h that codes for wavy hair are linked on the same chromosome;
4. Gamete that contained chromosome that contain allele B linked to allele h fuses with
gamete that contain chromosome that contain allele b linked to allele h;
[Total : 10]

N15P2 Q6

(a) Receptors for some hormones are found within their targets.
Explain why insulin receptors are found on the cell surface membranes of target cells
and never within the cells. [2]
1. Insulin is too large, cannot pass through any transient pores form within cell surface
membrane;
2. cannot pass through hydrophobic core of cell surface membrane because it is has
polar regions and will be repelled;

Comments: Do note that insulin receptors actually exist as linked dimers. Since
this question shows the 2 subunits dimerising, you need answer the question
according to the figure.
(b) Use Fig. 6.1 to explain how the presence of insulin is able to trigger a response inside
the target cell. [3]
1. Insulin has a specific 3D conformation* that is complementary* to the
extracellular ligand-binding site of the insulin receptor;
2. This ensures specificity in binding between insulin and the insulin receptors resulting
in the dimerization* of two receptor subunits;
3. Conformational change in the intracellular domain of receptor results in activation of
intrinsic tyrosine kinase;
4. Intrinsic tyrosine kinase activity of each subunit in the intracellular domain cross-
phosphorylates /autophosphorylates the tyrosine* residues on the other subunit;
5. Other relay proteins inside the target cell will be able to bind to the phosphorylated
tyrosine residues and become activated themselves;
This will enable the signal to be transduced within the cell until an appropriate cellular
response is reached.
 Raffles Institution Nov 2015 (H2 Biology) Paper 2 (for 9744 syllabus)

(c) Using Fig. 6.2., describe the main effects of insulin on different target cells. [6]
1. Binding of insulin to the insulin receptor will lead to cellular responses such as to trigger
the translocation of vesicles with glucose transporter-4 (GLUT4)* to the plasma
membrane of the target cells;
2. This will increase the number of glucose transporters (GLUT4) on the plasma membrane;
3. increasing the permeability of the plasma membrane to glucose. There will be an increase
in the uptake of glucose from the blood by these cells, causing blood glucose
concentration to drop;
4. The glucose taken up by the cells will be used to synthesize glycogen* via a series of
condensation reactions (i.e. glycogenesis);
5. Glycogen synthesis is catalyzed by glycogen synthase,
6. which is an enzyme activated as a result as the insulin signalling;
7. The glucose taken up by the cells will increase the rate of glycolysis and can also be
broken down by aerobic respiration to form intermediates (e.g. acetyl coA) which is then
used for fatty acid synthesis*;
8. This process is catalysed by various enzymes which are activated as a result of insulin
signalling.
[Total : 11 marks]
N15P2Q7
(a) State what is meant by the term, biological species. [2]
1. a group of organisms of the same species are capable of interbreeding* and;
producing fertile, viable offspring*;
2. are reproductively isolated from other species;

(b) Explain how new species arise. [5]

1. When a population is separated into 2 sub populations as they geographically
isolated* due to a physical barrier, interbreeding is prevented and gene flow is
disrupted*;
2. Different niches will present different selection pressures* and individuals best
adapted to the environment will have a selective advantage will be selected for and
favourable alleles will be passed on to the next generation and so the frequency of
favourable alleles will increase;
3. The 2 sub populations will evolve independently over time, accumulating different
mutations which will lead to changes to allele frequencies due to natural selection
and genetic drift;
4. Over hundreds and thousands of generations, across long periods of time,
accumulation of genetic differences led to each sub population becoming
reproductively isolated*;
5. Such that they can no longer interbreed* to produce viable, fertile* offspring and
hence new species are formed through allopatric speciation;

(c) Describe the advantages of using nucleotide sequences in reconstructing phylogenetic

relationships. [3]
1. They are objective. Molecular character states are unambiguous as A, C, G and T
are easily recognisable and cannot be confused;
2. Data is quantitative and easily converted to numerical form for statistical analysis.
The degree of relatedness can be inferred and quantified by calculating the
nucleotide differences between species;
3. Furthermore the mtDNA does not undergo recombination thus any changes to DNA
is due solely to the accumulation of mutations over time making it the ideal candidate
for a molecular clock. We can thus estimate the time of speciation;
 Raffles Institution Nov 2015 (H2 Biology) Paper 2 (for 9744 syllabus)

(d) Suggest, with reference to Fig.7.1 and Fig. 7.2, why breeding between C. lunulatus and C.
trifasciatus is possible. [3]
1. Since C. lunulatus and C. trifasciatus share a common ancestor (as seen in Fig 7.1);
2. and there are areas where they are found overlap (as seen in Fig.7.2);
3. they are able to interbreed and form hybrids;
[Total :13]

N15P2Q8
(a) Describe how DNA is arranged in the two structures. [4]
1. Eukaryotic DNA coils around proteins called histones to form nucleosomes;
2. followed by coiling around itself to form solenoid / 30nm chromatin fiber and
solenoid associates with scaffold proteins forming looped domains / 3000 nm
fiber;
3. Supercoiling of the loops to condense into the metaphase chromosome;
4. Eukaryotic DNA is linear and prokaryotic DNA is circular;
5. Prokaryotic DNA is associated with relatively fewer proteins (e.g. histone-like
proteins) to form loops and there is supercoiling to cause further compaction;

(b) State two ways in which the organisation of genes found in these two structures differ and
suggest one advantage of this to the bacterium. [3]
1. In bacteria functionally related genes are organised into an operon which
consists of an promoter, operator and structural genes while in eukaryotes
usually not organised into operon;
2. One promoter controls expression of more than one gene in bacteria while in
eukaryote one promoter controls expression of one gene;
Advantage of having operons in bacteria: [1 mark max]
3. Operons having related genes expressed under the control of one promoter
allows for fast response to environmental changes. This is important as bacteria
are unicellular and exposed to fluctuating environment.
4. Functionally related genes in an operon are expressed together as a set when
necessary for economical use of energy and resources;
[Total: 7]

N15P2Q9
(a) Describe the structure of a homologous pair of chromosomes at the start of [6]
meiosis
1. Homologous pair of chromosome consists of 2 homologues, one of paternal
and one of maternal origin;
2. Homologous chromosomes have the same size, shape, centromere
position* and staining pattern*;
3. The homologous chromosomes have the same genes at corresponding
loci*;
4. Homologous chromosomes are non-identical due to the presence of
different alleles;
5. Each homologue comprise of two sister chromatid* held together by a
centromere*;
6. Homologous chromosomes pair up to form bivalents/tetrads.
7. Each chromatid consist of a molecule of negatively charged*, DNA coiled
around 8 positively charged* proteins called histones*;
8. they form nucleosomes* that come together to form a solenoid;
9. which coils to form looped domains which further supercoil to form a short
and thick structure called chromatid.
 Raffles Institution Nov 2015 (H2 Biology) Paper 2 (for 9744 syllabus)

(b) Outline the behavior of chromosomes during meiosis [8]

1. At prophase I*, chromatin* coils, shortens and thickens into a condensed
chromosome;
2. homologous chromosomes* pair up by a process called synapsis* to
form a bivalents*;
3. Chiasmata* formation and crossing over* occurs;
4. such that exchange of equivalent portion of genetic material or alleles
occur between non-sister chromatids* of homologous chromosomes*;
5. At metaphase I*, homologous chromosomes* arranged as pairs at
metaphase plate/equator*;
6. At anaphase I*, each homologue is pulled by a shortening kinetochore
microtubule (that attaches to the centromere) towards opposite poles ;
7. At anaphase II*, centromeres* divide and sister chromatids separate to
form daughter chromosomes and;
8. are pulled by a shortening kinetochore microtube (that attaches to the
centromere) toward opposite poles;
9. At telophase II*, chromosomes reach the poles of the spindle where they
decondense and become diffuse/indistinct;

(c) Explain the role of nuclear envelope and centrioles during meiosis. [6]
Nuclear Envelope
1. Nuclear envelope is made of double membrane made up of phospholipid
bilayers;
2. During prophase I, nuclear envelope disintegrate* so that spindle fibres
can attach to the centromere* and the chromosomes can be pulled to
opposite poles;
3. During telophase II, nuclear envelope reforms around the chromosomes to
form the nucleus;
Centrioles
4. A pair* of cylindrical, rod-like structures;
5. which are perpendicular/right angle/ 90o* to each other;
6. Each consists of 9 triplets* of microtubules* arranged in a ring;
7. After the centrioles replicate during interphase, the two pairs of centrioles
move to opposite poles* during prophase*;
8. The centrioles become part of the microtubule-organising centre* in
which the assembly of spindle fibres and asters occur;
9. Spindle fibres are needed for the separation of chromosomes and
chromatids during meiosis I and II;

N15P2Q10
(a) Describe the role of NAD and FAD in cellular respiration. [6]

1. Organic molecules are oxidized during glycolysis, link reaction and Krebs
cycle* and the electrons (and protons) from the oxidation process are
transferred to the coenzymes NAD+* and FAD+* to form NADH* (reduced
NAD+) and FADH* (reduced FAD+) respectively;
2. NAD and FAD serves as mobile electron (and proton) carriers to carry the
high energy electrons and protons from these organic molecules to the
electron transport chain* on the cristae of mitochondria;
3. High energy electrons in NADH and FADH are used to reduce electron
carriers of the electron transport chain*, while NADH and FADH itself gets
re-oxidised;
4. As electrons pass down the chain, the release of energy in a series of redox
reactions is coupled to the phosphorylation of ADP to form ATP*;
5. Protons liberated in the oxidation of NADH and FADH is used to establish
the proton gradient* necessary for ATP synthesis (the H+ can be either
Raffles Institution Nov 2015 (H2 Biology) Paper 2 (for 9744 syllabus)

pumped into intermembrane space or combines with oxygen to form water

in the matrix).
6. Re-oxidation of NADH and FADH allows the regeneration of NAD+ and
FAD+*, allowing it to pick up more protons and electrons from Krebs cycle,
link reaction and glycolysis, so that these reactions can continue;
7. Each reduced NAD in the matrix yields 3 ATP;
8. and each reduced FAD in the matrix yields 2 ATP through oxidative
phosphorylation*;
9. During anaerobic respiration, re-oxidation of NADH allows the regeneration
of NAD+* (during alcohol or lactate fermentation) allowing glycolysis to
continue. [6 max]

(b) Outline the main stages of the Krebs cycle. [8]

1. Krebs cycle takes place in matrix of mitochondria and when oxygen is

present;
2. Acetyl CoA (2C) formed through link reaction combines with oxaloacetate
(4C) to form citrate (6C);
3. Citrate is decarboxylated and dehydrogenated to form α-ketoglutarate (5C)
and NADH.
4. Each decarboxylation step results in a loss of carbon in the form of a carbon
dioxide;
5. Regeneration of oxaloacetate (4C) involves one decarboxylation step and
three dehydrogenation steps to yield 2 NADH*, 1 FADH2* and 1 CO2;
6. Electrons (and protons) originally from glucose molecule have now been
transferred to electron carriers NAD+* and FAD*;
7. NAD+ + 2H+ + 2e- NADH + H+ (or reduced NAD)
8. FAD + 2H+ + 2e- FADH2 (or reduced FAD)
9. 1 ATP* is also produced through substrate-level phosphorylation during this
regeneration process;
10. All the carbon in glucose is lost as carbon dioxide;
11. Altogether 1 molecule of glucose will yield 6 NADH, 2 FADH2 & 2 ATP
through the Krebs cycle. The coenzymes with their reducing power will next
be transported to the electron transport chain where the bulk of ATP is
generated. [8 max]

(c) Explain how ATP is produced in anaerobic respiration. [6]

1. Anaerobic respiration takes place in the absence of oxygen;
2. Oxygen serves as the final electron acceptor* in electron transport
chain*;
3. Without oxygen both link reaction* and Krebs cycle* subsequently stop;
4. As NAD+* cannot be regenerated* from NADH* in mitochondrion;
5. Anaerobic respiration takes place in the cytoplasm of the cell via
glycolysis*;
6. with a net of 2 ATP molecules* produced via substrate-level
phosphorylation for each glucose molecule oxidised;
7. Pyruvate is reduced by pyruvate dehydrogenase to ethanol and carbon
dioxide with the regeneration of NAD+ in yeast;
8. In mammal NAD+ can be regenerated by converting pyruvate by lactate
dehydrogenase to lactate;
9. NAD+ regenerated in both processes ensures steady supply of NAD is used
for glycolysis to continue. [6 max]
[Total: 20]
 Raffles Institution Nov 2015 (H2 Biology) Paper 3 (for 9744 syllabus)

Nov 2015 H2 Biol Paper 3

N15P3Q1
The polymerase chain reaction (PCR) is a three-stage process.Describe what occurs in
(a) (i) The first stage at 95°C, [2]
1. heating to 95 C separates two strands of the DNA double helix;
2. by breaking the hydrogen bond between complementary bases through
increased molecular vibrations, denaturing the DNA;
(ii) the second stage. [2]
1. Cool to around 64 C, for DNA primers to anneal;
2. Primers anneal to complementary 3’ end of each template/single strand;
(b)
(i) Suggest and explain why Taq DNA polymerase is now obtained from genetically
modified E.coli. [2]
1. The use of genetically modified E.coli would allow for mass production of Taq
DNA polymerase;
2. Taq polymerase can be easily obtained;

(ii) With reference to your knowledge of PCR, explain why a half-life of 40 minutes at 95°C
allows many cycles of PCR before the enzyme needs to be replaced. [3]
A single cycle of PCR is very short/much shorter than 40 minutes;
1. Many cycles of PCR can occur within a single half-life of Taq polymerase;
2. As sufficient amounts of enzyme are present to carry out many cycles of PCR
before replacement is required;

(c) Explain how RFLP analysis of DNA samples from imported grain and non-GM grain can
show whether or not the imported grain has been genetically modified. [6]
1. Cut the DNA samples from the imported grain and non-GM grain with same
restriction enzyme* to obtain different-sized DNA fragments;
2. DNA is separated according to size in gel electrophoresis where negatively-
charged DNA* migrates towards the positive electrode/anode when subjected to
an electric field / current;
3. Meshwork of agarose fibres impedes movement of longer fragments more than
shorter fragments resulting in smallest fragments moving furthest/largest
fragments least far from well;
4. Carry out Southern blotting using radioactive probe* for VNTR/STR repeat
sequence followed by visualising the banding pattern using autoradiography / x-
ray film
5. Banding pattern in DNA fingerprint is due to different alleles/markers producing
different bands in gel resulting in the unique banding pattern for different DNA
samples;
6. Different bands arise due to polymorphic nature of different DNA samples, there
will be variations in number and location of restriction sites and number of
tandemly repeated nucleotide sequence among different DNA samples;
7. Genetic fingerprint of imported grain can be compared against fingerprint of non-
GM grain to see how closely related they are;
8. If fingerprint pattern is similar between the two different DNA samples, then
imported grain is non-GM;
[Total: 15 ]
N15P3Q2 (OUT OF SYLLABUS)
N15P3Q3 (OUT OF SYLLABUS)
 Raffles Institution Nov 2015 (H2 Biology) Paper 3 (for 9744 syllabus)

N15P3Q4
Planning Question
Suggested answer scheme:

Part 1: Aim
To investigate the time taken for the milk droplet of different concentrations to sink, and
hence determine the concentration of the milk provided by the supplier.

Part 2: Theory
(Main Theory): [T1: 1 mark for any 2 points from 1 to 5]

1. Milk is denser than the standard copper sulfate solution, and hence will sink to the bottom.
2. Density of the milk can be determined by the rate taken for the drop of milk to sink to the
bottom of standard copper sulfate solution, measured by the time taken (dependent /
measurable variable)
3. When milk is diluted with water, the density of the diluted milk will decrease and hence take
a longer time for the drop of milk to sink to the bottom. (predicted trend)
4. Milk of the same density will sink at the same rate.
5. Hence the density of the sample milk can be determined by comparing the rate it sinks to
the standard graph plotted using results of the rate of sinking of a set of standard
concentrations of milk.

Independent variable :
(at least 5 concentrations of milk solution at equal interval) AND description of how it is
controlled : 20%, 40%, 60%, 80% and 100% milk solution [2]

6. Prepare 10 cm3 20%, 40%, 60%, 80% and 100% milk solution by mixing appropriate
volumes of undiluted milk and distilled water in a test-tube as shown in the table below.
Use a 10cm3 syringe to measure the volume stated in the table.
Concentration of milk Volume of undiluted Volume of distilled Total volume
/% milk /cm3 water used /cm3 /cm3
20 2 8 10
40 4 6 10
60 6 4 10
80 8 2 10
100 10 0 10

Part 3: Procedure
1. Pilot Test
Conduct a pilot experiment to determine suitable range of independent variables used,
suitability of apparatus, and concentration of solution (e.g. copper sulfate solution). [P:
1 mark]
2. Annotated diagram
Set-up simple, diagram probably not needed.

3. Numbered steps in procedure

4. Fill a 100cm3 measuring cylinder with 100 cm3 of copper sulfate solution. [constant
variable]
5. Using the syringe with the attached needle, draw up 1cm3 of the 100% milk
6. Insert the tip of the syringe into the copper sulfate solution. Ensure the tip of the needle is
about 1cm into the copper sulfate solution. [constant variable]
7. In a controlled manner, slowly release 1 drop of the milk [constant variable] from the
syringe into the copper solution. Start the stopwatch.
Raffles Institution Nov 2015 (H2 Biology) Paper 3 (for 9744 syllabus)

8. Stop the stopwatch once the drop of milk reaches the bottom of the measuring cylinder.
9. Record the time taken for the drop of milk to reach the bottom of the measuring cylinder
[dependent variable] in the table below.
10. Calculate the rate taken for the drop of milk to sink using 1/time taken (s-1).
[Dependent Variable: 1 mark – steps 9 and 10, show how to obtain dependent
variable including calculation, method of obtaining must be scientifically sound]

11. Repeats steps 1 to 7 twice. These serve as replicates to check that no anomalies are
present. [R1: 1 mark for both replicates and repeats, including how they are carried
out and why they are carried out]
12. Repeats steps 1 to 8 using 20%, 40%, 60%, 80% milk solution, as well as the milk
sample from the supplier.
13. Repeat entire experiment (steps 1-9) twice more to check for reproducibility.
[R1: 1 mark for both replicates and repeats, including how they are carried out and
why they are carried out]

14. control
Keep all variables constant. Set up control experiment using water in place of milk.
This is to show that the presence of milk that causes the sinking of the droplet.
[Co: 1 mark for either control, including how it is carried out and reason why it is
performed]

Part 4: Data recording and processing:

Table showing rate of sinking of milk droplet
[T1: 1 mark for any full table including correct units – for time / rate &
concentrationeither pH or temperature]
Concentratio Time taken for drop of milk to sink / s Rate of sinking
n of milk /% Replicate 1 Replicate 2 Replicate 3 Average of drop of milk /
s-1
20
40
60
80
100
Sample milk
from supplier
 Raffles Institution Nov 2015 (H2 Biology) Paper 3 (for 9744 syllabus)

Standard graph of rate of sinking of milk droplet for different concentrations of milk

Rate of sinking
of drop of milk
/ s -1

20 40 60 80 100 Concentration
of milk / %

As shown above, use the standard curve to determine the concentration of the milk in the
sample from supplier.

Part 5 : Risks and precautions [1 mark]

(i) Be careful when using the sharp need, in order to prevent pricking oneself.
Wear gloves and goggle when measuring copper sulphate as they could be are irritants.
[Total: 12]

N15P3Q5 (OUT OF SYLLABUS)

Raffles Institution Nov 2016 (H2 Biology) Paper 2 (for 9744 syllabus)

Nov 2016 H2 Bio Paper 2

1 Fig. 1.1 shows the effect of increasing substrate concentration on the rate of an enzyme-catalysed
reaction in the presence and absence of a non-competitive inhibitor.
Competitive inhibitor

Fig. 1.1
(a) Explain why, in the reaction with the enzyme only, as substrate concentration increases:
(i) the rate of reaction increases at first [2]
1. As substrate concentration increases, frequency of effective collisions* between
enzyme and substrate molecules increases;
2. Rate of enzyme-substrate complex* formation increases and rate of reaction
increases as active sites* of enzymes are readily available and substrate
concentration is limiting;

(ii) the rate of reaction becomes constant. [2]

1. All active sites* of enzymes are saturated with substrate at any one point in time;
2. Concentration of substrate is no longer limiting and enzyme concentration is limiting,
rate of reaction will remain constant (graph plateaus).

(b) Explain why in Fig 1.1, the addition of a non-competitive inhibitor causes the reaction to become
constant at a lower rate. [2]
1. Inhibitor binds to site other than active site and changes conformation* of active site*;
2. Hence inhibitor effectively decreases availability of enzymes as it forms an enzyme-inhibitor
complex;
3. Effects of inhibition cannot be overcome by increasing substrate concentration;

(c) Draw, on Fig. 1.1, the approximate shape of the curve if a competitive inhibitor were added
to the enzyme instead of a non-competitive inhibitor. [2]
1. Correct shape and labelled;
2. approaches maximum rate of reaction;

(d) Suggest why the penicillin molecule is an effective inhibitor of transpeptidase. [2]
1. Cell wall peptide and penicillin are similar in conformation;
2. Penicillin can act as a competitive inhibitor*for the active site *of transpeptidase;
Hence, cell wall peptides cannot bind to active site and cross-links of cell wall peptides
cannot form.

[Total: 10]
Raffles Institution Nov 2016 (H2 Biology) Paper 2 (for 9744 syllabus)
2
(a) Identify the molecules labelled A and B on Fig. 2.1. [2]

A: messenger RNA
B: RNA polymerase (R: general transcription factor as the molecule is unzipping and
separating the double helix DNA)

(b) Explain how the control elements shown in Fig. 2.1 influence transcription. [4]
promoter: enhancer:

Promoter
1. serve as recognition site for the binding of general transcription factors* and RNA
polymerase*;
2. to initiate transcription*;
3. has critical elements, TATA box* that determines the precise location of
transcription start site;
4. has critical elements, CAAT and GC boxes* to improve efficiency of promoter by
recruiting general transcription factors* and RNA polymerase* to promoter.

Enhancer
5. when bound with specific transcription factors* known as activators*, promotes
assembly of transcription initiation complex*
6. when bound with specific transcription factors* known as activators*, may recruit
histone acetylase* and chromatin remodeling complexes* to decondense chromatin
(increase accessibility of promoter to general transcription factors and RNA polymerase)
7. increase frequency of transcription

(c) Describe the role of a silencer control element in transcription. [2]

1. allow binding of specific transcription factors* called repressors*

2. decreases the frequency of transcription of the genes they control by preventing
assembly of transcription initiation complex* at promoter / preventing the binding of
RNA polymerase* and general transcription factors* at promoter.

There are a number of differences between prokaryotic transcription and the eukaryotic
transcription shown in Fig. 2.1. [2]

(d) Describe a feature of the control of prokaryotic transcription that is not shown in Fig. 2.1. [2]

1. formation of RNA polymerase holoenzyme* which is made up of core RNA polymerase

and a sigma factor*;
2. sigma factor recognizes and binds to both the -10 sequence/Pribnow box* and -35
sequences
3. The more similar the -10 and -35 sequences are to the consensus sequences, the stronger
the promoter and therefore the higher frequency of transcription.
[Total: 10]
Raffles Institution Nov 2016 (H2 Biology) Paper 2 (for 9744 syllabus)
3
(a) Describe the changes shown in Fig. 3.1. [3]
1. Upon infection, the number of HIV viruses increase to a peak/maximum at week 3 and
decline steeply to near 0 at week 8.5;
2. From the week 9 to the year 6 (since primary infection), viruses remain relatively constant
near to 0 with 2 small peaks at year 2 and year 4;
3. From 6 to 8 years after infection, viruses increase steeply to a large number and plateau from
year 8 to 10;
4. The T helper cells decrease steeply from initial infection to week 3 and rises and remain
relatively constant till year 3;
5. From 4 to 9 years, the T helper cells decrease gradually to near to 0;
6. From 9 to 10 year, the T helper cells remain constant near to 0;

(b) Suggest how the changes in the number of T helper cells shown in Fig. 3.1 would affect the
health of an untreated HIV-infected individual over the course of the infection. [3]
1. T helper cells help to activate specific naïve B cells into plasma B cells to make antibodies for
antibody-mediated response; (CM I also added in underline)
2. the HIV infects more and more T helper cells, the levels of T helper cells lower from 4 to 9
years to near to 0, as the infected T cells are destroyed;
3. The increasing loss of T helper cells leads to impaired immune responses in the affected
individual who then becomes increasingly susceptible to opportunistic diseases;

(c) Why is HIV described as a retrovirus? [2]

1. A retrovirus is a RNA virus that duplicate via reverse transcription in the host cell;
2. using the RNA genome as a template, reverse transcriptase* produce DNA from its
RNA genome by complementary base pairing;

(d) Explain why viruses are described as obligate parasites. [2]

1. An obligate parasite is an organism that cannot live independently of its host;
2. They require the host cell to complete their life cycle and reproduce;

[Total: 10]
4
(a) Identify structures A, B, C and D, as shown on Fig. 4.1. [4]
A : mRNA
B : subunits of ribosomes
C : transport vesicle
D : membrane of the rough endoplasmic reticulum (RER)

(b) Use Fig. 4.1 to suggest how the structures labelled B attach to the endoplasmic reticulum
through the growing polypeptide. [2]
1. Molecules called signal recognition particle (SRP) bind to one end of growing polypeptide
at ribosome;
2.The molecules bind to receptor protein on membrane of B, to bring ribosomes to it;

(c) Suggest why the newly synthesised polypeptides shown in Fig. 4.1 cannot pass directly
into the cytosol. [1]
1. They have to be folded in correct conformation and undergo glycosylation in lumen of
RER before being passed to Golgi apparatus for further modification;

(d) Describe what happens to the newly synthesised polypeptides released from the rough
endoplasmic reticulum. [3]
1. Polypeptides are enclosed in a transport vesicle;
2. It fuses with cis/ convex face of Golgi apparatus* and undergoes chemical modification/
eg: glycosylation;
3. Golgi apparatus targets and sorts the polypeptides and secretory vesicles are released
Raffles Institution Nov 2016 (H2 Biology) Paper 2 (for 9744 syllabus)
from cis/ concave face;
4. The secretory vesicles fuse* with cell membrane and release polypeptides via
exocytosis;

[Total: 10]

5
(a) Draw a genetic diagram to explain the results of the first cross in which all offspring had
purple stems and green leaves.
Use the symbols N for the allele for purple stems, n for the allele for white stems, G for the allele
for green leaves and g for the allele for yellow leaves. [3]

Parental phenotype Purple stems, yellow leaves X White stems, green leaves
Parental genotype NNgg X nnGG

Correct parental phenotype + genotype [1]

Correct gametes [1]

Meiosis nG
Ng
Gametes X

Random fertilization

F1 genotype NnGg
F1 phenotype Purple stems, green leaves

Correct offspring phenotype + genotype [1]

(b) Explain the significance of the chi-squared value for these results. [4]

1. P > 0.90
2. Since p > 0.05, at a level of significance of 5% we do not reject the null hypothesis that
expected results are similar to the observed results;
3. The difference between the observed and expected phenotypic ratios is not significant
difference and is due to chance;
4. Inheritance of these genes follows Mendel’s laws of independent assortment, random
segregation and dominance.

(c) Suggest the advantages of using Fast Plants, instead of conventional crop varieties, as
sources of useful mutations that can then be introduced into broccoli and cabbage through
cross-breeding. [3]

1. Larger number of random new mutations that can result from fast plants (while in
cross-breeding, there is typically only reshuffling of combinations of alleles)
2. Fast plants have increased cycles of cell division, which could be due to mutations in
tumour suppressor genes
3. (Mutations in tumour suppressor genes means that) when there are further random
mutations that arise, cell cycle will not be halted for DNA repair resulting in
accumulation of further mutations
[Total: 10]
6 OUT OF SYLLABUS
Raffles Institution Nov 2016 (H2 Biology) Paper 2 (for 9744 syllabus)
7
(a) Suggest how N. diardi, of Borneo and Sumatra, evolved as a separate species from N.
nebulosa, found in the rest of Asia. [3]
1. When the ancestors of N. nebulosa, occupied new niches Sumatra and Borneo;
2. they were geographically isolated due to the presence of the sea from other sub-populations N.
nebulosa and could no longer interbreed with them and hence gene flow was disrupted*;
3. The different niches in Sumatra and Borneo presented different selection pressures* and so
individuals with favourable traits and hence a selective advantage were selected for, increasing
frequency of favourable alleles;
4. Over time the different sub populations N. nebulosa evolved independently of each other, their allele
frequencies changed as they accumulated different genetic mutations, and were subjected to genetic
drift and natural selection. Over a long period of time this led to reproductive isolation* and formation
of N. diardi, a new species (i.e. macroevolution) occurred;.

(b) There are a number of differences between populations of N. diardi on Borneo and
Sumatra. These are regarded as different sub-species.
Suggest why they are not regarded as separate species. [2]
1. They are not regarded as separate species as they are capable of interbreeding* and producing
fertile, viable offspring*;
2. and they are reproductively isolated* from other species;
3. usually have similar morphological, physiological and behavioural features;

(c) Explain whether the data in Fig. 7.2 provide sufficient evidence on their own for the
existence of two separate species of Neofelis. [2]
From the data, it can be seen that Neofelis diardi and Neofelis diardi
1. speciated about 1.4 million years ago from a common ancestor;
2. and have the same genus name but different species name/specific epithet, suggesting that they
are different species;
3. However, only by checking if they can interbreed and produce fertile viable offspring will there be
sufficient evidence that 2 separate species of Neofelis exist;

(d) Describe the advantages of using DNA sequence data in constructing phylogenies such as that shown in
Fig. 7.2. [3]
1. The DNA sequence data is objective*. Molecular character states are unambiguous* as A, C, G and T
are easily recognisable and cannot be confused;
2. The DNA sequence data is quantitative* and is easily converted to numerical form* and hence are
amenable to mathematical and statistical analysis and hence computation. The degree of relatedness
can be inferred and quantified by calculating the nucleotide differences between species;
3. The DNA sequence data can be used to compare species which are morphologically indistinguishable
due to convergent evolution* or are because they are very closely related;
[Total: 10]
8
(a) Describe what is happening in:
(i) phase 1 [2]
1. During carbon fixation, carbon dioxide* combines with RuBP*;
2. to form an unstable 6C compound that will immediately split to form 2 molecules of (3C) glycerate
phosphate* (GP);
3. Carbon fixation is catalyzed by enzyme, RuBP carboxylase* (Rubisco);

(ii) phase 2 [2]

1. NADPH is the reducing power used to reduce* GP to glyceraldehyde-3-phosphate (G3P);
2. ATP is the source of energy required;
3. Triose phosphate/G3P is the first sugar formed in photosynthesis and the end product of Calvin cycle;

(iii) phase 3 [2]

1. 5 molecules of G3P are used to regenerate* 3 RuBP so that the cycle of carbon dioxide fixation can
continue.
2. This requires 3 ATP;
Raffles Institution Nov 2016 (H2 Biology) Paper 2 (for 9744 syllabus)
(b) Light is not a factor that directly limits the rate of the Calvin cycle but it is needed for the
Calvin cycle to continue.
Outline why light is needed for the Calvin cycle to continue. [2]
1. The products from light reaction, ATP and NADPH are required for the reduction* of GP to G3P;
2. ATP is also used in the regeneration of RuBP*;

(c) State two environmental factors that can directly limit the rate of the Calvin cycle and explain
how they act. [2]
factor 1: Low carbon dioxide concentration (0.04%) in atmosphere
explanation: low carbon dioxide concentration decreases frequency of effective collisions*
between CO2, ribulose bisphosphate (RuBP) and rubisco thus, decrease enzyme-substrate
complex* formed per unit time, lowering rate of carbon fixation, which limits rate of Calvin cycle.

factor 2: Low temperatures (winter in temperate countries)

explanation: low temperature, leads to lower kinetic energy of CO2, ribulose bisphosphate
(RuBP) and rubisco, decreases frequency of effective collisions* between them thus, decrease
enzyme-substrate complex* formed per unit time, lowering rate of carbon fixation, which limits
rate of Calvin cycle.
[Total: 10]

9 (a) Describe the different functions of proteins in cell surface membranes. [7]
Transport proteins
1. Allow facilitated diffusion* of polar/charged* molecules or ions (solute) across the
membrane via transmembrane channels or carrier proteins;
2. Channel proteins are transmembrane proteins that have a hydrophilic pore for diffusion of
ions or polar molecules OR
3. Carrier proteins where hydrophilic molecules to bind to them / @ specific binding sites first
before they undergo a conformational change that results in transport of molecules across
membrane
4. For process of osmosis where water moves down a water potential gradient through
aquaporins of selectively permeable membrane;
5. Assist in active transport* of polar/charged molecules or ions across membrane via
protein pumps/ carrier, against concentration gradient using ATP*; e.g. Na-K pump are involved
in active transport;
6. For transport of molecules in large quantity against concentration gradient, receptor-
mediated endocytosis* occurs where specific ligands bind to receptor proteins on the
membrane causing invagination of membrane;
Cell to cell recognition & adhesion
7. Glycoproteins are involved in cell-cell recognition* to distinguish cells as ‘self’/’non-self’
as basis of immune system;
8. For cell-cell adhesion where cells of the same type to form tissues/ membrane protein of
adjacent cells may hook together to form various kinds of junctions such as tight junctions or gap
junctions
Receptor
9. As a receptor protein which a specific ligand will bind to.
10. The formation of ligand-receptor complex will initiate an intracellular signaling cascade for
signal transduction.
Cell membrane structure
11. Membrane proteins function to stabilise membrane structure as they are non-covalently
bound to cytoskeleton (on cytoplasmic side) & extracellular matrix (on extracellular side);
Enzyme
12. Some proteins which are enzymes that carry out catalytic/chemical reaction at cell
membrane e.g. adenylyl cyclase that catalyses conversion of ATP to CAMP/ acetylcholinesterase
on postsynaptic membrane
R : Act as electron carriers (e.g. cytochromes) which are part of the energy transfer systems that are
utilised during photosynthesis and respiration (question asked for cell surface membrane)
Raffles Institution Nov 2016 (H2 Biology) Paper 2 (for 9744 syllabus)
(b) Describe the process of endocytosis. [6]
Endocytosis
1. Is usually engaged to bring macromolecules or molecules in large quantity into the cell.
2. ATP* is needed for the rearrangement of microfilaments / microtubules
3. For macromolecules, there is phagocytosis* where pseudopodia* are formed and extended
outwards to engulf the macromolecule (large, insoluble).
4. When the ends of the pseudopodia fuse*, a vesicle* / vacuole containing the solid matter is pinched
off and enters into the cytoplasm;
5. There is also pinocytosis* which is to bring in tiny vesicle* of aqueous medium when a small area of
the plasma membrane invaginates
6. For transport of molecules in large quantity against concentration gradient, receptor-mediated*
endocytosis occurs where specific ligands bind to receptor proteins on the membrane causing
invagination of the membrane
(c) Outline the functions of membranes within cells. [7]
1. Enables compartmentalization of cell/ acts as a barrier to separate contents of cells from external
environment;
2. e.g. enzymes for Krebs cycle found only in matrix of mitochondrion/ hydrolytic enzymes are contained
in lysosomes and not in cytoplasm;
3. Controls exchange of substances across membrane between organelle and cytoplasm e.g. only
pyruvate can enter mitochondrion but not glucose; A: creation of proton gradient across
cristae/thylakoid membrane;
4. It contain enzymes for biochemical or metabolic reactions e.g. ATP synthase for phosphorylating ADP
to ATP;
5. Thylakoid membrane* of chloroplast* is site where photophosphorylation* takes place with
photosystems I and II are found with photosynthetic pigments;
6. It is site for ATP synthesis at inner membrane of mitochondrion* and thylakoid membrane of
chloroplast;
7. electron carriers in electron transport chain transport electrons down electron transport chain and
release energy required to pump protons;
8. from matrix to inner membrane space in mitochondria / from stroma to thylakoid space in
chloroplasts to create a proton gradient;
9. ATP synthase allows chemiosmosis to occur and hence production of ATP;
10. Highly folded cristae increases surface area to hold more ETC, ATP synthase for oxidative
phosphorylation/ stacks of thylakoid membranes in the form of granum to hold more photosystems,
ETC and ATP synthase for photophosphorylation;
11. Site for protein synthesis at rough endoplasmic reticulum* where translation takes place at
ribosomes at RER membrane;
[Total: 20]
10 (a) Explain how different types of bonding hold protein molecules in shape. [7]
1. Hydrogen bond*;
2. Oxygen (e.g. O of C=O group*) and nitrogen (e.g. N of -NH group*) are electronegative ( -).
Hydrogen of -NH or -OH group is electropositive ( +).Electropositive and electronegative atoms form
hydrogen bonds;
3. Ionic bond*;
4. Formed between oppositely-charged* R groups* of amino acids;
5. Hydrophobic interaction*;
6. Formed between non-polar* R groups* which are hydrophobic;
7. Disulfide bond* / bridge
8. Formed only between two cysteine amino acids by oxidation of sulfydryl (-SH) groups, which
contains sulphur*;
9. In collagen, covalent cross-links* form between lysine* residues at C and N ends of
adjacent/parallel tropocollagen molecules;
Examples of shapes:
10. Secondary structure*: α-helix or β-pleated sheet held in place by hydrogen bonds between C=O
and N-H groups of polypeptide backbone;
11. Tertiary structure*: single polypeptide chain further extensively folded and bended into specific
Raffles Institution Nov 2016 (H2 Biology) Paper 2 (for 9744 syllabus)
conformation held in place by hydrophobic interactions, ionic bonds, disulfide bridges and
hydrogen bonds formed between R groups;
12. Quaternary structure*: association of two or more polypeptide chains held together by same four
types of interactions involved in tertiary structure;
(b) Describe how amino acids are joined together. [6]
Note: If diagram is drawn, it should be properly annotated.

1. Amino acids undergo condensation* to form polypeptides with removal of 1 water molecule;
2. OH group from carboxyl group and H atom from amino group contribute to formation of water molecule
(showing OH and H in diagram coming together);
3. Box up and label peptide bond*;
4. Correct structure and label of amino acids;
5. Box up and label amino terminus and carboxyl terminus;
6. Polypeptide formed from condensation;
7. This reaction is catalysed* by enzyme, peptidyl transferase*, at ribosomes*;

(c) Outline the structure of haemoglobin and relate this to its function. [7]
1. 4 polypeptide subunits: 2 -globin* subunits and 2 -globin* subunits;
2. each subunit is arranged so that most of hydrophilic amino acid side chains are on external surface
while hydrophobic amino acid side chains are buried in interior;
3. makes it soluble in water/aqueous environment, can be transported and carry O2 from lungs to tissues
vice versa;
4. each subunit is made of globin polypeptide and a prosthetic (non-protein) component called haem
group*;
5. each haem group consists of a porphyrin ring* and an iron ion (Fe2+)*;
6. Fe2+ of haem group binds temporarily to O2, so 1 Hb molecule can carry up to 4 O2, at a time forming
oxyhaemoglobin; (ref. transport oxygen in blood)
7. 4 subunits held together by intermolecular interactions formed between R groups (hydrogen bonds,
ionic bonds and hydrophobic interactions), allows movement that influences affinity for oxygen
allowing for cooperative binding* of oxygen;
8. As a result binding of one oxygen molecule to one haemoglobin subunit induces a conformational
change in remaining 3 subunits so that affinity for oxygen increases;
[Total: 20]
Raffles Institution Nov 2016 (H2 Biology) Paper 3 (for 9744 syllabus)

Nov 2016 H2 Bio Paper 3

Answer all questions

1 (a) Different restriction enzymes (endonucleases) such as Hindlll and EcoRI are naturally
produced by particular species of bacteria.
Describe the natural functions of these restriction enzymes. [2]
1. Used as a defense mechanism by bacteria against bacteriophage by cutting up foreign DNA,
hence restricts multiplication of viruses;
2. Enzyme that recognizes and bind a specific 4-6 base pair DNA sequence called a restriction
site as its active site is complementary to the DNA sequence;
3. Enzyme cuts/breaks phosphodiester bonds* on specific positions on both DNA strands;
(b)
(i) Use the information in Table 1.1 to compare the effect of pH on immobilised P1 nuclease
and free P1 nuclease. [4]
1. Compare peak + quote data
The optimum pH for immobilized P1 nuclease was higher at pH 6.0, with nuclease
activity of 68 arbitrary units, whereas free P1 nuclease peaked at pH 5.5, with
nuclease activity of 65 arbitrary units.
2. Immobilised P1 nuclease activity was lower than free P1 nuclease from pH 5 to pH5.5.
+ quote data from either pH
e.g. immoblised P1 nuclease activity was 49 a.u. whereas free P1 nuclease was 56
a.u. at pH 5.0, or immoblised p1 nuclease activity was 7 a.u. lower than free P1
nuclease at pH 5.0
3. Immobilised P1 nuclease activity was higher than free P1 nuclease from pH6.0 to
pH7.0. + quote data from any 1 pH
e.g. immoblised P1 nuclease activity was 49 a.u. whereas free P1 nuclease was 56
a.u. at pH 5.0, or immoblised p1 nuclease activity was 7 a.u. lower than free P1
nuclease at pH 5.0);
4. Both had similar trend of increasing in activity from pH4.5 to pH5.5/6.0 then decreasing
from pH5.5/6.0 to pH7.0 + quote data

(ii) With reference to your knowledge of enzyme structure and the information in (b),
explain the higher activity of immobilised P1 nuclease, compared to free P1 nuclease, at
temperatures above 30 °C, as shown in Table 1.2. [3]
1. At higher temperatures, intramolecular vibrations increase, which results in the
breaking of bonds that determine the conformation of the enzyme in the free P1
nuclease.
2. Resulting in the denaturation of the enzyme due to a change in conformation of the
enzyme active site, decreasing the rate of enzyme activity of free P1 nuclease
3. Binding of P1 nuclease to matrix, stabilize the enzyme’s structure at higher
temperatures, reducing intramolecular vibrations, prevents breaking of bonds, and
retaining the conformation of enzyme active site.
4. Activity of free P1 nuclease starts to drop at 60°C which is lower than that of 70°C in
immobolised P1 nuclease.
(c)
(i) Not in syllabus
(ii) Suggest why P1 nuclease was needed in the production of this particular recombinant DNA
molecule. [3]
1. P1 nuclease was needed to break down the different sticky ends produced by the
digestion of HindIII and EcoRI.
2. As it is able to break down single-stranded DNA, leaving the double stranded DNA
fragment intact.
3. Scientist will then be able to use the double stranded DNA fragment to synthesize
complementary stick ends/add ligase to join the desired DNA fragments directly to
form a recombinant DNA molecule.
Raffles Institution Nov 2016 (H2 Biology) Paper 3 (for 9744 syllabus)
[Total: 14]

2 In humans and other animals, the protein collagen is found in many regions of the body, including
bones, tendons, muscles and skin.
The COL1A1 gene codes for one of the polypeptides in a particular type of collagen.
(a) From your knowledge of collagen structure, explain why the length of any exon is divisible
by 9. [3]
1. Collagen has a repeating amino acid sequence of glycine-X-Y/three amino acids;
2. Where each amino acid is deterimined by a codon of three nucleotides on the mRNA,
3. Thus, three of such codons will be 9 nucleotides in the exon.

(b) Not in syllabus

(c) Not in syllabus

[Total: 13]

3 (a) The genetic disease X-linked SCID (severe combined immunodeficiency) can be treated by
bone marrow transplants. This process can be carried out very early in a child's life.
(i) The bone marrow contains blood stem cells that are described as being multipotent.
State what is meant by multipotent. [1]
1. Multipotent stem cell has ability to divide* and differentiate* to form limited range of
cell types / differentiate into red and white blood cells.;

(ii) Describe the characteristics that identify all stem cells. [2]
1. A stem cell is an undifferentiated/unspecialized* cell capable of undergoing
proliferation* and self-renewal*;
2. and retains potential to differentiate* to produce specialized cells upon receiving
appropriate molecular signals*;

(b) Not in syllabus

(c)
(i) Suggest how the mitochondrial genome can code for about 40 genes when it is only four times
the size of the SCID gene. [2]
1. As mitochondrial genome are similar to prokaryotic genome, they are much smaller, as
it has lesser non-coding sequences compared to genes in the chromosomes.
2. SCID genes are large due to introns* which are sequences that are spliced out during
post-transcriptional modifications.
(ii) Not in syllabus
[Total: 13]

4 Planning question

Penicillin is an antibiotic that is used to treat infections by killing bacteria.

In an investigation, a student was provided with Petri dishes containing nutrient agar. Each Petri
dish had already been inoculated with Staphylococcus sp. and incubated to produce an evenly
distributed growth of the bacteria (a bacterial lawn).
In a trial investigation, the student cut four wells into the agar and added distilled water to one of
them and three different concentrations of penicillin solution to the others.
Fig. 4.1 shows the result after incubating the Petri dish for 24 hours.
Raffles Institution Nov 2016 (H2 Biology) Paper 3 (for 9744 syllabus)

The clear zones around the wells containing penicillin solution showed that the Staphylococcus sp.
had been killed, whilst none had been killed by the distilled water. The student noticed that the two
highest concentrations of penicillin tested had clear zones that were the same size.
The student read that garlic has long been used to treat wound infections. During the Second
World War, the Russian army ran out of penicillin and used garlic as an antibiotic, leading to garlic
being called the Russian penicillin.
The student wanted to find the most effective concentration of garlic solution. This is the lowest
concentration of garlic solution that gives the largest clear zone possible.
Using this information and your own knowledge, design an experiment to find the lowest
concentration of garlic solution that gives the largest clear zone possible.

You must use

100 cm3 20% garlic solution,
200 cm3 distilled water,
prepared 100 mm diameter agar plates, with a lawn of Staphylococcus sp.,
disinfectant (sterilising) solution and paper towels.

You may select from the following apparatus:

set of cork borers with diameters from 4 mm to 15 mm,
normal laboratory glassware, e.g. beakers, measuring cylinders, graduated pipettes,
glass
rods, etc.,
incubator,
autoclave (a pressurised oven for heat sterilising apparatus and materials),
Bunsen burner,
sticky tape,
mm ruler,
syringes.

Your plan should:

have a clear and helpful structure such that the method you use is able to be repeated by
anyone
reading it,
be illustrated by relevant diagrams, if necessary ,
identify the independent and dependent variables,
describe the method with the scientific reasoning used to decide the method so that the
results,
are as accurate and reliable as possible,
show how you will record your results and the proposed layout of results tables and graphs,
use the correct technical and scientific terms,
include reference to safety measures to minimise any risks associated with the
Raffles Institution Nov 2016 (H2 Biology) Paper 3 (for 9744 syllabus)
proposed experiment.
[Total: 12]
Main theory Effectiveness of active compounds in garlic solution against Staphylococcus
can be determined by placing garlic solutions of different concentrations onto
wells on prepared 100 mm diameter agar plates, with a lawn of
Staphylococcus sp.*.
Garlic solution will diffuse through agar and inhibit growth of bacteria.
Measurable This results in a clear zone around the well that has no bacteria growth. This
quantity is known as zone of inhibition and its diameter is a measure of effectiveness
of garlic solution against bacteria.
Trend The higher the concentration of garlic, the more it will diffuse across agar,
hence a zone of inhibition with a larger diameter due to more bacteria dying.
Most effective concentration of garlic solution is lowest concentration of
garlic solution that gives largest zone of inhibition. Further increase in
concentration of garlic solution does not increase diameter anymore.
Independent 20% garlic solution* diluted at 4 concentrations using syringes and sterile
variable distilled water* should be prepared first, under aseptic techniques:
Concentration of garlic Volume of stock garlic Volume of distilled
solution (%) solution (ml) water* (ml)
20 10.0 0.0
16 8.0 2.0
12 6.0 4.0
8 4.0 6.0
4 2.0 8.0
Dependent Growth of bacteria, as indicated by diameter of zone of clearance (mm)
variable measured using a mm ruler.
Controlled variables
1) Volume of (how) using same cork borer eg: 10 mm diameter, puncture 6 wells of same
garlic solution size on given Petri dish with agar
poured into (why) wells need to be same size so that constant volume of garlic solution
wells on agar will be placed in the wells as amount of active compounds in garlic solution
plate affects bacterial growth
2) Temperature (how) incubator set at 37 C for 12 hours
where bacteria (why) temperature needs to be kept constant as bacteria growth is affected
is incubated by temperature
Replicates (how) 3 replicates are the 3 different petri dishes, each with 5 wells with
different concentrations of garlic solution
(why) check for anomalous results
Repeats (how) repeat entire experiment 2 more times
(why) check for reproducibility
Negative control (what) set up control experiment by filling up well with sterile distilled water
on bacteria lawn. All other variables and procedures should remain
constant.
(why) control shows no clear zone, where bacteria growth is not inhibited.
Bacterial growth in other wells is due to presence of garlic solution.
Raffles Institution Nov 2016 (H2 Biology) Paper 3 (for 9744 syllabus)
Annotated
Zone of inhibition 100 mm diameter agar
diagram plates containing lawn of
measured in mm. This is
Staphylococcus bacteria
measure after incubating
4% garlic 8% garlic
agar plates 37 C for 12 solution solution
hours.

12% garlic
solution

20% garlic
solution
Zone of inhibition
measured in mm. This is
measure after incubating 16% garlic
solution
agar plates 37 C for 12
hours.
Note: In diagram, 16% and 20% garlic solutions produce zones of same diameter.
That means 16% garlic solution is minimum concentration.

Fig. 1 - Set up of experiment to find

Results Concentration of Diameter of zone of inhibition (mm) Average

garlic solution (%) Replicate 1 Replicate 2 Replicate 3 diameter (mm)
20
16
12
8
4

Average diameter
of zone of inhibition
(mm) Lowest concentration of
garlic solution that produces
largest zone of inhibition

Concentration of
garlic solution (%)
Risks and Experiment needs to be carried out under sterile microbiological techniques to
precautions prevent contaminating bacteria from growing on agar plates. Before any work is
started, bench must be wiped with disinfectant (sterilizing) solution and
paper towels.*

Additional precautions (name 1):

e.g.: cork borer need to be sterilized using autoclave
e.g.: when placing garlic solutions into wells on agar plate, pipette garlic solution
using micropipette tips that have been sterilized using autoclave. It is
important to work near a lighted up Bunsen burner
e.g.: water need to be sterilized using autoclave
Raffles Institution Nov 2016 (H2 Biology) Paper 3 (for 9744 syllabus)
2. As bacteria may be harmful, gloves must be used when handling agar plates.
Used materials including agar plates need to be properly disposed of by
sterilizing using autoclave.
Improvement Lowest concentration of garlic solution – experiment can be further refined
to find exact concentration of garlic solution. Repeat experiment under same
conditions, use smaller intervals
e.g.: If minimal concentration is 16%, repeat with 15.0,15.5%, 16.5%,
17.0%, 17.5% garlic solution.

5 Free-response question
(a) Explain what is meant by restriction fragment length polymorphism (RFLP). [5]
1. Restriction Fragment Length Polymorphism (RFLP) refers to the unique banding pattern among
individuals when the genomic DNA are digested by restriction enzymes separated by gel
electrophoresis;
2. e.g 1. single nucleotide polymorphisms (SNPs, pronounced ‘snips’) / differences at a single base-pair
in DNA sequences in homologous regions;
3. due to mutations;
4. can result in variations in number and/or location of restriction sites;
5. Due to e.g 2. variations in the number of tandem repeats;
6. e.g. 1 or 2 produce fragments of different length among individuals called restriction fragment length
polymorphisms (RFLPs);
7. Upon visualization and analysis, a unique banding pattern is produced for each individual;

(b) Outline the process of nucleic acid hybridisation. [10]

1. To detect the RFLP pattern of a gene, the DNA sample is digested with restriction enzyme and;
2. Then the digested DNA samples are separated using gel electrophoresis;
3. ds DNA is denatured / made single-stranded and by alkaline / NaOH solution;
4. and transferred to a nitrocellulose membrane;
5. at exactly the same position as they were in the gel;
6. The nitrocellulose membrane incubated with a radioactive single stranded DNA probe*, that is
complementary in sequence* to part of the target sequence / gene;
7. DNA fragments containing this part of the target sequence will hybridise to the probe by
complementary base-pairing*;
8. After hybridisation, membrane is washed to remove any excess unhybridised probes.
9. Using autoradiography/ by placing X-ray film* over the membrane, the banding pattern can be
visualised;
10. The radioactivity of the bound probes exposes the film to form an image corresponding to the
bands that have base-paired to the probe;

(c) Explain how RFLP analysis facilitates the process of DNA fingerprinting. [5]
1. When the genomes from 2 individuals A and B, are cut using the same restriction enzyme (e.g.
BamHI);
2. If the 2 individuals differ in the number of tandem repeats (of the sequence ATTAC) at a particular
locus on particular chromosome;
3. The individual with the larger number of tandem repeats present, (e.g. individual B) generates a
longer restriction fragment which travels a shorter distance down the gel during electrophoresis;
4. while the individual with fewer number of tandem repeats present, (e.g. individual A) generates a
shorter restriction fragment which travels a longer distance down the gel during electrophoresis;
5. The 2 different-sized fragments are detected using the same probe which has a sequence
complementary to segments of both fragments e.g. ATTAC.
6. Will result in fragments of different lengths being produced;
7. Can be discriminating enough to generate a DNA fingerprint / unique genetic profile for each
individual.
[Total: 20]
 2017 H2 A level Paper 2

1
(a) (i)
• 2

(ii)
• Protein / polypeptide

(iii)
• binds / holds 2 glycogen molecules together;

(b)

1. Large molecule/long chain so insoluble in water ;

2. Hence does not affect water potential of cells / no osmotic effect ;

3. Helical molecule so more glucose units per unit volume, thus compact structure for storage ;

4. Extensively branched so provides many ends for hydrolysis of glycosidic bonds ;

5. which releases more glucose molecules per unit time ;

(c)

Cellulose Glycogen
monomer β-glucose* α-glucose*
bonds β-1,4 glycosidic bonds* α-1,4 and α-1,6 glycosidic
bonds*
Orientation of Alternate glucose units are All glucose units in the chain have
monomer rotated/inverted 180o with same orientation
respect to each other
Structure of each forming straight / linear forms extensively branched helical
molecule unbranched chains ; coils ;
Bonds between Hydroxyl groups* projecting No interchain hydrogen
molecules outwards in both directions allow bonding* in glycogen
intermolecular/ interchain
hydrogen bonding *
à leading to microfibril
formation for cellulose
Note: must be written in full sentences

2
(a) (i)
1. Glucose is polar* and thus, hydrophilic*;
2. Glucose is too large to pass through the transient pores of the phospholipid
bilayer ;
3. The hydrophobic core* of the phospholipid bilayer would repel glucose ;
 4. Transport proteins / protein channel provides a hydrophilic* channel/pore through
the membrane for the passage of the solutes;

(ii)
1. The protein is a transmembrane protein that;
2. has amino acid residues with R groups that is non-polar / uncharged, allowing it to
form hydrophobic interactions with the hydrophobic core of the phospholipid
bilayer;
3. The protein has a hydrophilic pore/channel that is lined with amino acids with polar
/ charged R groups that allows hydrophilic glucose molecules to pass through the
membrane;

(iii)
• Q

(b)
Describe:
1. Increase in external concentration of glucose increases the rate of glucose uptake
2. When external concentration of glucose increase from 0 - 1 mmol dm-3, rate of glucose
uptake increases sharply from 0 – 180 arbitrary units;
3. When external concentration of glucose increases from 1 – 11 mmol dm-3, rate of glucose
uptake increases from 180 – 395 arbitrary units, but rate of glucose uptake increase less
with each unit increase in external glucose concentration;
4. Rate of glucose uptake plateaus at 395 arbitrary units when external glucose
concentration increase from 11 - 12 mmol dm-3;

Explain:
5. Increase in external glucose concentration increases concentration gradient of glucose
across the membrane;
6. Maximum rate of glucose uptake is reached when all glucose transporters/transport
proteins are saturated;

3
(a)
Haemoglobin:
1. Haemoglobin is a globular protein;
2. It has a quaternary structure and comprises 4 polypeptide chains/subunits/2α and 2β
chains;
3. Each subunit is associated with a haem* prosthetic group;

Collagen:
4. Collagen is a fibrous protein;
5. It has a quaternary structure and comprises 3 polypeptide chains coiled around each
other;
6. Each polypeptide is a loose helix;
(b)
Haemoglobin
Molecular structure Function
1. Each subunit is bound to a haem* Allows for each subunit to bind to an oxygen
prosthetic group; molecule;
2. Hemoglobin has a quaternary structure Having multiple subunits allow for the protein
as it is made up of 2α and 2β subunits; to exhibit cooperativity in its ability to bind to
oxygen. Binding of 1 oxygen molecule causes
a conformational change that allows
subsequent oxygen molecule to bind more
easily;
3. Amino acids with hydrophilic R groups Allows molecule to be soluble in water so that
are found on the exterior of the protein it can be easily transported in blood
and amino acids with hydrophobic R
groups are buried in the interior of the
protein;

Collagen:
Molecular structure Function
1. Numerous hydrogen bonds between Gives rise to high tensile strength*
OH groups on different polypeptides /
loose helix
2. Staggered arrangement of collagen Minimises point of weakness;
molecules within the fibrils
4

Direction of Direction of
movement movement

Lagging Leading
strand strand

Fig. 4.1

(a) On diagram

(b)
J DNA polymerase
K DNA ligase

(c) 1. This is because DNA polymerase* works only in the 5’ to 3’ direction;

2. Hence, strand extension occurs in the direction away from the replication fork / strand extends
in opposite direction with respect to the replication fork;
3. Synthesis of the new strand can only occur in fragments as the DNA double helix unzips;

5
(a) 1. The virus infected cells reproduced to produce precancerous cells that possess nuclei with
non-integrated viral DNA;
2. The virus infected cells become larger and go through more rounds of cell division, such that
the layer of cells no long have a distinct boundary;
3. As the virus reproduces in the host cell, more infectious virus particles are released to infect
even more cells;
4. Over time, the viral DNA can be intergrated into the host cell genome such that that cells are
permanently altered genetically;
5. The cells in the basal layer are no longer cuboidal, but are irregular shapted;
6. They also continue to show uncontrolled cell proliferation and penetrate the basal membrane
and the dermis to result in invasive cancer;
(b) 1. p53 gene is a tumor suppressor gene coding for the p53 protein;
E6 protein destroys the p53 protein and hence:
2. Prevents apoptosis/DNA repair;
3. This allow for uncontrolled cell division;
4. And accumulation of mutations;

(c)
1. In most other cancer, p53 gene would go through loss-of-function mutation* to produce
non-functional p53 protein;
2. In cervival cancer, p53 gene is not mutated and functional p53 protein can be produced
3. The infection with human papillomavirus introduces the E6 gene into the cells allowing for the
production of E6 protein which destroys the p53 protein, hence the function of the p53 protein
is disrupted, even though the protein is functional;

6
(a)
• (recessive) epistasis;

(b)
(i)
aaBB, aaBb and aabb (must state all. R: aa_ _)

(ii)
AAbb and Aabb (must state all)

(iii)
AABB, AaBB, AABb and AaBb (must state all. R: A_B_)

(c)
(i)
Parental phenotype: Black x Agouti
Parental genotype: Aabb x aaBB
Gametes: Ab ab aB

Offspring genotype: AaBb aaBb;

Offspring phenotype: 1 agouti: 1 albino;

(ii)
Parental phenotype: Black x Black
Parental genotype: AAbb x Aabb;
Gametes: Ab ab Ab ab

Offspring genotype: AAbb Aabb Aabb aabb

Offspring phenotype: 3 black : 1 albino

7
(a)
(i)
NADH

(ii)
Glycolysis, Link reaction, Kreb Cycle

(iii)
NADH donates electrons to the electron transport chain and is re-oxidised to form NAD;

(b)
1. Electrons passed down electron carriers in order of electronegativity releases energy
which is used to pump H+ from the matrix to the intermembrane space;
2. This produces a proton motive force / H+ concentration gradient, which allows H+ to
diffuse through ATP synthase* back to the matrix;
3. This diffusion of H+ through ATP synthase is coupled to the phosphorylation of ADP to
form ATP by ATP synthase*;

(c)
1. Molecule B is water;
2. Oxygen serves as the final electon acceptor* of the electron transport chain;
3. And accepts electrons and protons to form water;

(d)
photophosphorylation oxidative phosphorylation
Electron donors Water is the electron donor in NADH and FADH2 are the
the non-cyclic pathway while electron donors to the first
Photosystem I is the electron electron carrier of ETC;
donor in the cyclic pathway;
Electron acceptor NADP+ is the final electron Oxygen is the final electron
acceptor in the non-cyclic acceptor (and it combines
pathway while Photosystem I with H+) and is reduced to
is the final electron acceptor water;
in the cyclic pathway;
Electron transport chain 2 electon transport chains 1 electron transport chain;
involved;
Pathway of electrons Linear for non-cyclic and Linear;
circular for cyclic;

8
(a)
F Ligand
G Receptor
(b)
1. Different cells have different intracellular signaling proteins;
2. Hence different signaling pathways are activated to produce different responses;

(c)
Each protein kinase is activated by phosphorylation;

(d)
1. Each step results in an increase in the number of activated molecules;
2. For example active protein kinase 1 is able to phosphorylate and activate many protein
kinase 2, which can go on to activate even more protein kinase 3;

(e)
1. Mutations in genes could be gain-in-function mutations* which would code for proteins
that have an increased function;
2. For example, gene coding for protein kinase 1 could be mutated such that protein kinase
1 is always active, without requiring the activated relay molecule;
3. This could result in cells dividing excessively, without presence of appropriate signals,
causing uncontrolled cell division;
4. Alternatively, mutations could also be loss-of-function mutations*, coding for proteins
which are non-functional;
5. For example, gene coding for protein kinase 1 could be mutated such that protein kinase
1 cannot activate protein kinase 2;
6. This could result in cells being unable to trigger apoptosis or DNA repair, resulting in
uncontrolled cell division;

9
(a) (i) 1. As island size increases, the total number of plant species also increases;
2. Eg. 1000 species on island 200 000 km2 to 2500 species on islands 800 000 km2
3. Some exceptions/anomalies exist: island of 75 000 km2 has same numebr of
species (8000) as island of 100 000 km2 or smaller island 150 000km2 has 1200
species, which is more than a larger island of 200 000 km2 with 1000 species;

(ii)
Larger/Bigger islands have (any 2)
1. greater no. of habitats;
2. Greater variety of habitats/environments/named example;
3. More available niches;
More unique environments for selection to occur and hence more species;

(b) 1. Islands are geographically isolated* as they are surrounded by water that acts as a
physical barrier preventing interbreeding. This results in the disruption of gene flow*;
2. The islands due to their differing habitats / environments, present many niches for species
to fill – idea of adaptive radiation;
3. e.g. of differences. Soil type – sandy or clayey, availability of water, availability of shade,
plant types;
4. Thus (common) ancestral species on different islands were exposed to different
selection pressures* and natural selection act on them;
5. There exist variation in population and those with favourable traits have a selective
 advantage to the local conditions and will be selected for, increasing frequency of
favourable alleles / and will survive, reproduce and pass on their alleles to the next
generation;
6. As different sub populations evolved independently of each other, their allele frequencies
changed as they accumulated different genetic mutations, and were subjected to genetic
drift and natural selection.
7. Over a hundreds and thousands of generations, each population on the different islands
became reproductively isolated*;

10
(a)
1. IgG has an antigen binding site/variable region that is;
2. Complementary in shape and charge to the epitope on the antigen of the virus, allowing it
to recognise and bind to the antigen;

(b)
1. Binding of IgG to the antigen causes the neutralization of the antigen and
2. prevents the antigen/virus from binding to the host cell surface receptor on the host cell
membrane and
3. prevents receptor mediated endocytosis from occurring;
4. Binding of IgG to the antigen also allows for opsonisation*, recruiting macrophages /
phagocytes via the Fc region;
5. increasing the frequency of phagocytosis* by macrophages/phagocytes

11
(a)
1. From mid1994 to the end 1997, there was a gradual increase in the percentage of coral
cover from about 42 to 50%;
2. From the end 1997 to mid 1998, there was a sharp decrease in the percentage of coral
cover from 50% to about 10%;
3. From mid 1998 to mid 2010, there was a gradual increase in coral cover from about 10%
to 45%;

(b)
This is because there were more algae growing on the dead coral à more food for herbivorous
fish.

(c)
Fish eat the algae on the corals and allow corals to recover.
Long-structured Q - 2017 A level P3Q1

1 (a) (i) 1. Disease is a health / physiological impairment as a result of;

2. Mutation* in the DNA;
3. which could have occurred in the individual with the disease, or inherited from
parents;
4. The mutation can be in the form of a nucleotide substitution, insertion or deletion;
5. which would cause a change in the mRNA codon and subsequently, a change in
the amino acid in the protein resulting in a loss-of-function;
6. Or could have resulted in a premature stop codon, producing a truncated protein;
7. Mutations could also be at the chromosomal level, where there is a change in the
number of chromosomes or chromosomal translocation, duplication, deletion or
inversion;

(ii) 1. The PKU phenotype would only be observed in individuals that are homozygous
recessive for the disease allele (genotype);
2. and when phenylalanine is present in the diet of the individual (environment);

(iii) Both molecules have

1. an amino group (NH2/NH3+);
2. a carboxyl group (COOH/COO-)

Importance to function:
3. to participate in condensation reaction;
4. to form peptide bonds in the production of proteins;

(b) (i) 1. Extract genomic DNA from cells obtained in the blood / from a mouth swab;
2. Carry out polymerase chain reaction (PCR)* using primers* complementary to
regions flanking the PKU allele;
3. Cut the amplified fragment using restriction enzyme*;
4. Carry out gel electrophoresis* to separate the fragments according to size,
5. Carry out Southern blot* and use radioactive probes complementary to the PKU
allele to detect specific fragments
6. Carry out RFLP analysis: compare with restriction patterns for known genotypes to
determine unknown genotype;

A: after PCR – carry out DNA sequencing & compare with known DNA sequences of
the normal allele and the PKU allele

(ii) 1. The results of the test would be a false negative, showing wrongly that the baby
does not have PKU;
2. No colony / reduced colony diameter would be observed;
3. Despite high levels of phenylalanine in baby’s blood, which should have allowed
for bacterial growth;
4. Bacteria growth was inhibited / bacteria was killed by the antibiotics present in the
blood;

(iii) 1. -2-thienylalanine competes with phenylalanine for the active site* of;
2. Aminoacyl-tRNA synthetase*, that joins phenylalanine* to its tRNA;
3. -2-thienylalanine is incorporated into polypeptides instead of phenylalanine,
resulting in a change in the primary structure / amino acid sequence;
 4. Tertiary structure / 3D conformation of the protein is thus changed;
5. producing non-functional bacterial proteins which resulted in bacterial growth being
inhibitied;

(c) 1. Loss of function of enzyme Q stops conversion of phenylalanine* to tyrosine*;

Less tyrosine results in:

2. less thyroxine* which leads to poor brain development;
3. less melanin* which causes fair skin and hair;
4. less dopamine* which results in poor transmission of nerve impulses;

(d) (i) P(PKU) = P(Carrier parent 1) X P(Carrier parent 2) X P(child affected)

=1/140 x 1/140 x ¼ = 1/ 78400 or one in 78 400

(ii) The frequency of PKU in cold and wet, Northern Europe is the highest in the world where 1 in10
000 are sufferers while sub-Saharan Africa which is hot and dry is lower by 10 times; (Quote data)
1. This is because fungi thrive in cold and wet climates in North Europe but not in sub-
Saharan Africa which is hot and dry. This means increased likelihood of renal cancer
in North Europe as the fungi produces, ochratoxin A; (link climate to fungi to cancer)
2. Selection pressure is ochratoxin A;
3. Homozygotes for the normal PKU alleles are selected against in North Europe as they
are not protected against ochratoxin A;
4. Homozygotes of PKU alleles are selected against because they have metabolic
deficiencies that result in poor intellectual and behavioural development;
5. When both selection pressures are applied, as happens in North Europe, the
heterozygotes are selected for as they are resistant to ochratoxin A while they do not
suffer the developmental deficiencies of PKU;
6. This is called heterozygote advantage*, a form of balancing selection where both
alleles are selected for under specific circumstances found in North Europe but not
found in Africa;
7. Heterozygotes are therefore able to survive and pass on the PKU allele to the next
generation, resulting in an increase in the frequency of the PKU allele over time;

[Total: 30]

2 (a) 1. Blood stem cells are unspecialized cells which are multipotent*; (Reject- toti/ pluri)
2. Ability to self-renew* and proliferate*
3. to maintain a constant pool of stem cells;
4. Ability to differentiate* into specialized cells in the blood, eg red blood cell, lymphocytes
(name 1 eg)
5. to replace dead cells that died;

(b) 1. The named T lymphocytes are T helper cells / lymphocytes;

2. A particular naïve T cell* will have T cell receptors (TCR) that can specifically recognise
the peptide of peptide:MHC complex on the antigen presenting cell (APC;
3. The APC secretes cytokines that will activate the naïve T cells which will undergo clonal
expansion and differentiation to form effector and memory T cells ;
4. T helper cells* secrete cytokines that stimulate/activate specific naïve B cells* to
become antibody-secreting plasma cells*;
5. Memory T cells* when re-exposed to the same pathogen/antigen, will recognize it and
mount a faster and stronger secondary immune response;
 (c) 1. Unforeseen circumstances from T cells lacking CCR5 / immune response might be
affected if T cells do not have CCR5 receptor;
2. Unintended changes in DNA as a result of gene editing may contribute to development of
cancer;
3. Blood stem cells are multipotent and hence cannot form a whole organism, hence no
debate of whether or not a life is “sacrificed”;
4. Potential to cure patients completely so patients no longer have to be on lifetime anti-viral
therapy;
5. AVP eg. Accessibility to treatment

[Total: 10]

3 (a) 1. During carbon fixation stage, CO2 is combined with ribulose bisphosphate (RuBP)*,
catalysing this reaction is RuBP carboxylase (RUBISCO);
2. to form an unstable 6 carbon molecule which;splits up immediately into 2 molecules of
glycerate phosphate* (GP);
3. NADPH is the reducing power used to reduce* GP to glyceraldehyde-3-phosphate* (G3P),
and ATP provides a source of energy;
4. ATP* and NADPH* are products from the light-dependent reaction;

(b) 1. At temperatures above the optimum range, enzymes involved in leaf and stem growth
denature*,
2. At temperatures below the optimum, rate of enzymatic reactions decreases;
3. At lower temperature, enzyme and substrate molecules have lower kinetic energy*
resulting in lower frequency of formation of enzyme-substrate complex*;

(c) 1. Soybean, Glycine max;

2. With an increase of 2oC, maximum temperature can reach 37oC which is above the
temperature at which crop fails for maize (35oC);
3. Mean temperature will rise to 29oC which is within the range for optimum growth of leaf
and stem (25-36oC);
4. Temperature range for optimum seed production is higher in soybean as compared to
maize, hence soybean would be able to produce more seeds at a higher mean
temperature / Quote data;
[Total: 10]

4 (a) Part I: Describe reproductive cycle of enveloped virus with reference to influenza
1. Virus attaches to host cell:
a. Influenza virus attaches to epithelial cells of the respiratory tract;
b. Hemagglutinin* binds to complementary sialic acid* receptors on the host cell;
2. Virus enters by endocytosis*: host cell membrane invaginates* and the virus is placed
in an endocytic vesicle / endosome;
3. Endosome fuses with a lysosome*, resulting in a lowering of pH;
4. Lowered pH causes viral envelope to fuse* with the vesicle membrane, releasing the
nucleocapsid* into the cytosol;
5. Capsid proteins are degraded by enzymes and the 8 segments of viral RNA genome
enter the nucleus;
6. In the nucleus: RNA-dependent RNA polymerase* uses the RNA genome as a
template* to synthesis a complementary RNA strand;
7. Complementary RNA strand moves into the cytosol and serves as the mRNA;
 8. Which is used as a template for translation* to synthesise viral structural components;
a. Capsid proteins are synthesised by free ribosomes* in the cytosol;
b. Glycoproteins are synthesised by ribosomes* at the rough endoplasmic
reticulum* and are eventually embedded into the host cell membrane;
9. Complementary RNA strand also used as a template* for synthesis of new viral genome
by RNA-dependent RNA polymerase;
10. Assembly of new viral progeny: Capsid proteins associate with glycoproteins at the cell
membrane, nucleoproteins associate with RNA genome and then interact with the capsid
proteins;
11. The host cell membrane evaginates* and the newly formed viruses leave the host cell by
budding;
12. acquiring the host cell membrane with the embedded viral glycoproteins;
13. Neuraminidase* facilitates the release of the viruses by cleaving the sialic acid residues
from the host cell receptor;

Part II: Why is this a reproductive cycle and not a life cycle
14. Not considered a life cycle as virus is not living;
15. Features of virus that causes it to be classified as non-living:
- Absence of metabolic activity eg. No respiration;
- No growth
- No movement
- Acellular

QWC: Both Part I and Part II addressed

(b) 1. Global warming is an increase in atmospheric temperatures around the world;

2. Increased temperature decreases survival of virus and hence lowers human exposure to
virus;
Seasonal pattern;
3. With global warming, there are more extreme weather events with summer being hotter
and winters being colder;
4. Survival of virus decreases in the hotter summer months due to higher temperature; ORA
5. Colder winters results in an increase incidences of viral diseases such as influenza in
winter months; ORA
Geographical pattern:
6. Temperature decreases with increased latitude;
7. Higher incidence of viral diseases around the equator / at lower latitudes vs higher
latitudes;

Ref to viral diseases involving a vector, eg. Dengue fever:

8. Mosquito vector, Aedes aegypti, lives and breeds in moist tropical regions;
9. With global warming, the duration for development/life cycle of mosquito from first instar
to emergence is shorter;
10. As an increase in temperature increases their metabolic processes by increasing rates of
enzyme-catalysed reactions;
11. resulting in faster development rate and decreasing the length of reproductive cycles and
stimulating hatching of eggs;
12. More mosquitoes result in greater transmission of dengue fever;
13. Increased temperature also results in shorter extrinsic incubation period as virus can
reproduce faster in mosquito vector;

Seasonal pattern:
14. Hotter summer months results in an increase incidence of dengue fever in summer;
 Geographical pattern:
15. Global warming results in temperate regions becoming warmer, such that conditions
become more suited for the survival of Aedes mosquitoes;
16. Mosquitoes will thus move to higher latitude so they spread to new areas expanding their
distribution;
17. Global warming also results in higher altitudes becoming warmer, thus mosquitoes will
be able to colonise altitudes higher up the mountain;

QWC: to address both changes in seasonal and geographical patterns with regards to viral
diseases, citing specific examples.

5 (a) Feature Eukaryotic cell Prokaryotic cell Method

(bacteria)
Cell size Larger: 10-100µm Smaller: 0.5 - 5µm in diameter
Light/electorn
in diameter microscopy
Nucleus Nucleus with No true nucleus / Light/electron
nuclear envelope No nuclear microscopy
present; envelope
Genetic Linear DNA Circular DNA Restriction digest and
material associated with associated with few electrophoresis
many proteins; histone-like
Found in proteins;
membrane bound Found in a region of
nucleus; the cytoplasm
No plasmids known as the
nucleoid region;
Plasmids present
Ribosome 80S; 70S; Electron microscopy
for protein Ribosomes may No ER present.
synthesis be attached to ER Ribosomes free in
or free in cytoplasm.
cytoplasm
Organelles Many; Few; Electron microscopy /
Many membrane No membrane Differential centrifugation
bound organelles bound organelles;
present;
Cell walls Composed of Composed of Gram staining for
cellulose in plants peptidoglycan or prokaryotes
& chitin in fungi murein
 Flagella Complex: Simple: Electron microscopy /
9+2 arrangement Lacking immunofluoresence
of microtubules; microtubules - just
Intracellular a single strand of
(enclosed by protein;
plasma Extracellular (not
membrane); enclosed by plasma
200 nm in membrane);
diameter 20 nm in diameter

QWC: link method to feature to be distinguished

(b) Part I: Expected problems

Prokaryotes in the human microbiome are
1. Small and therefore difficult to distinguish morphologically;
2. similar in structure in terms of shape or cell wall or function and therefore difficult to
classify;
(For example they may use oxygen the same way (aerobic or anaerobic) or obtain energy the same
way (autotrophs or heterotrophs) and hence will be difficult to distinguish between them and classify them ;

3. reproduce asexually/undergo conjugation/transduction and therefore difficult to

identify/distinguish
4. some may be difficult to grow culture in the lab and hence difficult to study
5. defining species in bacteria is difficult if we use biological concept of species as bacteria
divide asexually by binary fission

Part II: Advantages of using molecular methods

1. They can be used to compare all organisms which share common genes.
2. They can be used to compare organisms that are morphologically indistinguishable
due to convergent evolution or because they are closely related.
3. Molecular methods are objective as molecular character states are unambiguous (e.g.
A, C, G & T) whereas some morphological characters, such as those based on the shape
of a structure or colour, can be less easy to distinguish objectively.
4. They are quantitative as molecular data can be converted into numerical form and
statistical analysis performed to determine degree of relatedness by calculating
nucleotide differences between organisms.
5. Changes in nucleotide sequences accumulate over time with clockwork regularity
and this forms the basis of the molecular clock. We can thus estimate the time of
speciation of modern to ancient species.
6. Some molecular differences may not be reflected as a morphological difference while
small genetic differences may not result in a major phenotypic difference. This means
that molecular data does not underestimate nor exaggerate differences unlike
morphological analysis.
7. Molecular methods offer a large set of characters to be studied quickly. Each
nucleotide position can be considered a character to distinguish between species.
8. Nucleotide sequences for a rapidly increasing number of genomes & amino acid
sequences for many proteins can be accessed from electronic databases for
comparative study & classification of all life.
9. Specimens need not be complete or alive for comparative analysis so long as the
molecules survive degradation.
 2018 H2 A level Paper 2
Answer all questions.

1(a) Discuss how the arrangement of molecules in Fig. 1.1 supports the fluid mosaic model of the cell
membrane. [3]
1. It is referred to as ‘fluid” because the cell membrane comprises of phospholipids* and a G-
protein linked receptor* which are free to move laterally within a layer;
2. Presence of cholesterol* molecules within each phospholipid layer increases the fluidity of the
membrane;
3. It is referred to as ‘mosaic” because the random arrangement of the proteins embedded
amongst the phospholipid molecules resemble a mosaic pattern;

(b) Explain how the molecular structure of the protein shown in Fig. 1.1 enables it to function as a G-
protein linked receptor. [3]
1. G protein linked receptor (GPCR) a seven pass transmembrane protein consisting of 7 α-
helices* connected by three intracellular and three extracellular peptide loops.
2. The intracellular domain/cytoplasmic side of GPCR has a G protein binding site* that allows
binding of a heterotrimeric G protein complex.
3. The extracellular loops has a ligand binding site* at which a specific* signaling molecule can
bind to the GPCR.
4. When a ligand binds to the ligand binding site at the extracellular side of a GPCR it causes a
conformational change* of the intracellular domain /at the cytoplasmic side of the GPCR,
5. The activated GPCR can then activate an associated G protein by exchanging its bound GDP
for a GTP.

(c) A large range of stimuli can trigger the activation of G-proteins through G-protein linked receptors.
These stimuli can include light, calcium ions and hormones such as glucagon.

State the common effect that these diverse stimuli have on G-protein linked receptors. [1]
1. The stimuli can trigger a conformational change* in the intracellular domain /at the
cytoplasmic side of the GPCR, activating the receptor.

(d) Explain how the activation of the G-protein by the binding of glucagon to the G-protein linked
receptor triggers downstream signalling pathways that result in a cascade of enzyme-catalysed
reactions. [3]
1. When glucagon binds a G-protein linked receptor (GPCR), the GPCR will undergo a
conformation change in intracellular domain allowing G-protein to bind to it; and
2. G-protein is activated when it displaces its attached GDP for GTP;
3. Activated G-protein will translocate along membrane and bind to enzyme adenylyl
cyclase and phosphorylate it, thus activating it;
4. Adenylyl cyclase will catalyze conversion of ATP to cAMP, which binds to and activates
protein kinase A (PKA);
5. Activation of PKA will initiate a sequential activation of kinases resulting in a
(phosphorylation cascade) to eventually activate glycogen phosphorylase for
breakdown of glycogen;

[Total: 10]
2(a) Describe, with reference to Fig. 2.1, the effect of temperature on the rate of protein digestion by
thermitase. [3]
1. As temperature increases from 10°C to 76°C, the rate of protein digestion by thermitase
increases from 0 arbitrary units(a.u) to 650 arbitrary units(a.u).
2. The optimum temperature of the enzyme thermitase is 76°C where there is maximum rate of
protein digestion.
3. Further increase in temperature from 76°C to 90°C will cause the rate of protein digestion
decreases sharply from 650 a.u to 355 a.u.

(b) Explain the effect on thermitase of increasing the temperature above 80°C. [3]
1. As the temperature increases above 80°C, there will be greater (intramolecular) vibrations/
thermal agitation of the enzyme thermitase,
2. which breaks hydrogen, ionic bonds and other weak interactions that stabilizes the 3D
conformation resulting in denaturation*
3. The enzyme active site* no longer complementary in shape* and charge to the substrate
and
4. the rate of reaction decreases steeply from a rate of 570 a.u. to 340 a.u.

(c) Modified subtilisin is similar to subtilisin, but has had eight of its amino acids replaced with different
amino acids.

Describe and explain the effect of this modification on the activity of subtilisin. [4]
Describe
1. Modified subtilisin has a higher rate of protein digestion across all temperatures compared to
subtilisin.
or
2. Modified subtilisin has a wider range of temperatures in can work in, from 10°C to 90°C
compared to subtilisin which works from 10°C to 74°C.

3. Modified subtilisin has an optimum temperature of 76°C compared to subtilisin which has an
optimum temperature of 59°C.
4. The maximum rate of protein digestion in the modified subtilisin is 4 times higher than in
subtilisin (i.e. 319 a.u for modified subtilisin and 78 a.u for subtilisin).

Explain
5. The replacement of eight amino acids in the modified subtilisin has resulted in a more
thermostable enzyme.
6. The R groups* of these amino acids are able to form stronger bonds, such as strong covalent
bonds like disulfide linkages between cysteine residues, which can only be broken at higher
temperatures.
7. Hence, the conformation of the active site remains stable and is able to catalyse the reaction
longer and at higher temperatures.

[Total: 10]
3(a) Define each of the following types of stem cell and name a naturally occurring example of each.

(i) totipotent [2]

1. They have the ability to differentiate* into all cell types that make up an organism
including extra-embryonic tissue* such as placenta*, and hence are able to form
entire organism;
2. e.g. zygotic stem cells/ fertilised egg up to 8 cell stage/first three divisions;

(ii) pluripotent [2]

1. They have the ability to differentiate* into all of the cell types that make up an
organism except extraembryonic tissue* such as placenta* but cannot form the entire
organism alone;
2. e.g. inner cell mass of blastocyst*

(iii) multipotent [2]

1. They have the ability to differentiate* into only a limited and related range of cell types
and tissues in an organism to replace dead cells that died;
2. e.g blood/haematopoietic stem cells in bone marrow

(b) Induced pluripotent stem cells (iPSCs) are a type of pluripotent stem cell that are produced directly
from adult cells. To produce iPSCs, four genes need to be introduced into adult cells.
These genes code for transcription factors.

Explain why these transcription factors are necessary for the production of iPSCs from adult cells.
[2]
1. These transcription factors initiate the transcription of specific inactive genes in the
differentiated somatic adult cells;
2. and altered the gene expression patterns of the cells such that it was similar to that of embryonic
stem cells;
resulting in the production of iPSCs

3. These transcription factors such as repressors/activators* bind to the silencers/enhancers*;

4. initiate the transcription of inactive genes in the differentiated somatic adult cells;
5. and altered the gene expression patterns of the cells such that it was similar to that of embryonic
stem cells;
resulting in the production of iPSCs

A: upregulate proto oncogene (see below for example)/inhibit tumour suppressor gene.

1. One of these transcriptions factors is an activator that binds to the enhancer of a proto
oncogene
2. That upregulates transcription leading to increased cell proliferation.
3. Example is a protein that promotes cell division e.g. cyclin, growth factors.
(c) Explain how the use of iPSCs may overcome some of the ethical concerns of using other types of
stem cells in medical research and treatment. [2]
1. As iPS cells are not from an embryo,
A. there will be no destruction of embryos and hence not regarded as killing a life due to
some views that embryos are viable living organisms;
B. Some object to extracting stem cells from an embryo to make replacement body cells is
treating embryo as just a source of spare parts;
C. Once human status is denied to embryos, this precedent may extend to other categories
of human beings such as profoundly disabled or elderly infirm;
2. Since the patient’s own iPSCs can be used, there will be no foreign tissues or antigens
introduced into the patient and thus will not result in immune response/tissue rejection and
thus there will be no need for immunosuppressant drugs;

[Total: 10]

4(a) With reference to structures shown in Fig. 4.1, explain how an enveloped virus such as influenza:

(i) enters a host cell [4]

1. Haemagglutinin* binds to sialic acid receptor* on host cell membrane;
2. Virus then enters host cell by endocytosis* where host cell membrane invaginates*
and pinches off, placing virus in an endocytotic vesicle;
3. Vesicle then fuses* with a lysosome*, and the resulting drop in pH stimulates fusion of
viral envelope with vesicle membrane;
4. Releasing nucleocapsid* and eventually viral RNA into cytosol;

(ii) exits a host cell [2]

1. Capsid proteins associate with the nucleoprotein (viral genome and proteins) near host
plasma membrane;
2. Each new virus buds from the host cell;
3. Acquiring viral envelope from the host plasma membrane which has neuraminidase and
haemagglutinin embedded;

(b) With reference to Fig. 4.2, explain how antigenic shift occurs. [2]

1. Antigenic shift occurred where the RNA segments from the 2 different influenza virus packaged
into the same virion, giving rise to new combinations of RNA segments resulting in new
combination of glycoproteins in the novel virus;
2. Either
In 1957, H2N2 bird virus and H1N1 human virus infects same cell giving rise to new H2N2
human virus with new combination of RNA segments;

Or
In 1968, H3 bird virus and H2N2 human virus infects same cell giving rise to new H3N2 human
virus with new combination of RNA segments;
(c) Viruses such as influenza rapidly accumulate gene mutations.
Explain the advantage of this to the virus. [2]
1. Rapid mutations results in antigens like haemmaglutinin and neuraminidase having a different
conformation;
2. Thus cannot be recognized by memory* T and B cells of the immune system immune cells;
3. So it takes a longer time to remove the virus from the body allowing them opportunity to
replicate;
[Total: 10]

5 (a) Describe two ways in which the lac operon is similar to the trp operon. [2]
1. Both consist of several genes under the control of a single promoter
2. Both have an operator region for the binding of an active repressor that prevent transcription
3. The genes code for enzymes involve in the same metabolic pathway or functionally related
proteins are synthesised as a unit;
4. Each operon when expressed produce a polycistronic mRNA that has several start codons and
stop codons

(b) Describe four ways in which the lac operon is different from the trp operon. [4]
Lac opreon Trp operon
Type of operon Inducible operon that is switched on Repressible operon that is switched
in the presence of an inducer off in the presence of excess end
lactose product tryptophan
Effector Inducer is lactose which is the Corepressor tryptophan, an end
molecule substrate product
Repressor Active form Inactive form
synthesized in
Condition Allolactose binds to and inactivates Tryptophan binds to and activates the
affecting the repressor such that it cannot repressor such that it can bind to the
repressor bind to the operator operator
Type of Enzymes produced involved in a Enzymes produced involved in an
metabolic catabolic process of breaking down anabolic process of synthesizing
pathway lactose tryptophan
Default operon Operon is switched off by default Operon is switched on by default until
expression until the presence of lactose tryptophan is in excess
No. of genes/ lac operon codes for 3 different trp operon codes for 5 different
proteins coded enzymes enzymes

(c) Suggest the advantages to bacteria of arranging some genes in operons. [4]
1. Operon contain group of genes under the control of the same promoter that allow for functionally
related proteins to be synthesised as a unit;
2. An operon can be turned ‘on’ or ‘off’ according to certain changes/conditions so as to respond
rapidly and appropriately to the environment
3. Bacterium only produces enzymes when required allowing the bacteria to make economical
use of energy and resources/conserve resources i.e. relevant genes are expressed only when
necessary;
4. Especially since bacteria are able to use a variety of metabolites e.g. glucose is metabolised
preferentially over lactose, thus not economical to produce lac genes in the presence of
glucose;
5. The abovementioned provide a selective advantage to such bacteria
[Total: 10]
6(a) During the PCR cycle shown in Fig. 6.1, three different temperatures are required.

Explain why each of these three different temperatures is required during one PCR cycle. [3]
Note: Quote temperatures from the graph not the ones you memorise!

1. Heating to 95 C* causes the hydrogen bonds* between complementary bases of each strand
to break, causing denaturation of double-stranded DNA into single-stranded DNA and exposes
the bases for complementary base pairing;
2. Lower temperature of 55 C* allows primers* to anneal* specifically to the regions flanking the
target DNA sequence via complementary base pairing*, providing a free 3’-OH group for
chain extension;
3. Subsequent temperature of 70 C* is the optimum temperature of Taq polymerase* which
performs the chain extension/ elongation* by synthesising the complementary DNA strand
from the 3’ end of the primer;

BUT if question was a 6 mark question, please elaborate:

95 C
1. Heating to 95 C causes the weak hydrogen bonds* between complementary bases of each
strand to break due to increased molecular vibrations;
2. The denaturation of double-stranded DNA into single-stranded DNA exposes the bases for
complementary base pairing;

55 C
3. Lower temperature of 55 C allows primers* to anneal* specifically to the regions flanking the
target DNA sequence via complementary base pairing*;
4. The primers determine the segment to be amplified and provides a free 3’-OH group for chain
extension;

70 C
5. Is the optimum temperature of Taq polymerase* which performs the synthesis of
complementary* DNA strand;
6. Chain extension/ elongation* occurs from 3’ end of primer which provides free 3’ OH required
by polymerase;
(b) Explain the role of primers in the polymerase chain reaction. [2]
1. Primers are short, single stranded DNA sequences that can anneal to the target DNA sequences;
2. they are complementary* to the regions flanking the gene of interest and thus determine the
segment to be amplified;
3. they provide a free 3’-OH group for chain extension by DNA polymerase so that the gene of
interest can be amplified;

Teacher’s comment:
Many students did not mention point 1. It is useful to take note that when describing the role, it is
good to briefly mention the key structural feature.
For point 2, many students wrote that the primers bind to the 3’ ends of single-stranded DNA but
this statement is incorrect because a single-stranded DNA strand can comprise several genes but
primers bind to the 3’ ends of the target gene only.

For example,
INCORRECT х

5’ --------------------------------------------------------------------------------------------------------------------3’
Gene 1 Gene 2 Gene 3
3’ --------------------------------------------------------------------------------------------------------------------5’

CORRECT to specifically amplify Gene 2 √

5’ --------------------------------------------------------------------------------------------------------------------3’
Gene 1 Gene 2 Gene 3
3’ --------------------------------------------------------------------------------------------------------------------5’

For point 3, most students were able to score the marks for this. However, those who failed to
mention that the free 3’OH was needed by the DNA/ Taq polymerase were not awarded the
marks.

(c) PCR products are usually separated by gel electrophoresis.

Outline the main principles that allow gel electrophoresis to separate DNA fragments. [3]
1. Dense loading buffer is mixed with DNA sample to help it sink to the bottom of wells located
nearest the negative electrode/cathode;
2. Negatively-charged* DNA migrates out of well towards direction of positive electrode/anode
when subjected to an electric field / current;
3. Buffers contain ions which allows conduction of electric current;
4. Meshwork of agarose polysaccharides impede movement of longer fragments more than
shorter fragments;
5. causing them to migrate slower than shorter fragments and end up nearer to the well;

(d) Explain why marker DNA was included in this experiment. [2]
1. The marker DNA consists of a mixture of DNA fragments of known sizes;
2. it acts as a standard that the fragments of unknown sizes from the 3 DNA samples can be
compared with to estimate their fragment size;
[Total: 10]
7(a) Explain the term epistasis in this context. [3]
(Colourless precursor Yellow fruit Green fruit)

1. Gene A/a and B/b interacts to determine one trait, i.e. colour of squash
2. Genotype aa results in production of yellow pigment which is the intermediate required by
genotype bb to produce green fruit
3. Genotype A_ results in production of inhibitor that prevents colourless precursor to be converted
to yellow pigment
4. Genotype B_ results in production of inhibitor that prevents yellow pigment to be converted to
green pigment

(b) Draw a genetic diagram to explain the results of crossing the F1 offspring to produce the expected
12:3:1 phenotypic ratio in the F2 generation.

Use the symbols A, a and B, b to represent the alleles. [4]

Let A represent the dominant allele for production of inhibitor that prevents colourless precursor to
be converted to yellow intermediate.
Let a represent the absence of inhibitor.

Let B represent the dominant allele for production of inhibitor that prevents yellow intermediate to
be converted to green pigment.
Let b represent the absence of inhibitor.
or
Let A be the dominant allele for coloured and a be the recessive allele for white
Let B be the dominant allele for yellow and b be the recessive allele for green
F1 phenotype White X White
F1 genotype AaBb AaBb
Fertilisation to produce
F2

Gametes
AB Ab aB ab

AABB AABb AaBB AaBb

AB
white white white white
AABb AAbb AaBb Aabb
Ab
white white white White
AaBB AaBb aaBB aaBb
aB
white white yellow Yellow
AaBb Aabb aaBb aabb
ab
white white yellow green
 F2 genotypes 9 A_B_ : 3 A_bb : 3 aaB_ : 1 aabb

F2 phenotypic ratio 12 white : 3 yellow : 1 green

(c) Explain how the chi-squared test value of 2.88 supports dominant epistasis as the correct
explanation of these results. [3]
1. Since 0.10 < p < 0.25;
2. Therefore p > 0.05 at 5% level of significance, we do not reject the null hypothesis that there is
no significant difference between the expected and observed phenotypes
3. Difference between observed and expected results is not significant and due to chance,
therefore supporting dominant epistasis as the correct explanation;
[Total: 10]

8(a) Describe the changes in the percentage contributions to muscle respiration of fatty acids from blood
and glycogen in muscle during this long-distance run, as shown in Table 8.1. [3]
1. The percentage contribution to muscle respiration by fatty acids from blood was 37% for the
first 90 minutes and started to increase* significantly to 62% at 240 minutes;
2. The percentage contribution to muscle respiration by glycogen in muscle was highest at 36%
at 40 minutes but reduced* until it reached 8% at 240 minutes;
3. The percentage contribution from fatty acids and glycogen was similar in the first 40 minutes
but as the run progresses, the percentage contribution started to diverge with a lower
contribution from glycogen and a correspondingly higher contribution from fatty acids;
(comparison)

(b) During the long-distance run, ATP may be generated at times using the type of respiration that
normally occurs under anaerobic conditions. However, the athlete cannot use this type of
respiration continuously throughout the whole period of the long-distance run.

Suggest why this type of respiration cannot be used continuously by the athlete to generate ATP
during the long-distance run. [2]
1. Under anaerobic conditions, lactic acid fermentation* occurs;
2. Lactic acid accumulates in the muscles resulting in muscle fatigue;

(c) Explain the different patterns of change in the percentage contributions to muscle respiration of
glucose from food and glycogen in muscle during the long-distance run, as shown in Table 8.1. [4]
1. Over the duration of the run, the percentage contribution to muscle respiration from glycogen
in muscle decreases over time because glycogen, a carbohydrate store in the muscle is
progressively used up; data not required as mark was already allocated in part (a)
2. There was a significant increase to muscle respiration by glucose from food from 27% to 41%
at the 90 minute mark; Quote for glucose
3. This delay in increase could be a result of time taken to digest the food/absorption of glucose;
4. From the 90 minute onwards, the contribution by glucose from food gradually decreases to 30%
(as less food is available to generate glucose/fatty acids becomes a major energy contributor);
5. The changes in pattern is dependent on the energy sources available over time. Glycogen is a
ready energy source that is used first while glucose from digestion of food is used next;

(d) Suggest why fatty acids are not used more in the earlier stages of the long-distance run. [1]
It takes time to convert fat into energy. Lipase will need to breakdown fat to release fatty acids
followed by oxidation of fatty acids to give rise to acetyl-CoA;
[Total: 10]
9(a) With reference to Fig. 9.1, explain why the large ground finch, medium ground finch and small
ground finch are considered to be three separate species. [3]
1. The 3 different finches have their own distinct body masses and beak length in their
populations. The large ground finch population has body mass between 32 -39g and beak
length between 36-42mm, the medium ground finch population has a body mass between
18-24g and beak length between 24-28mm and the small ground finch population has a
body mass between 10-15g and beak length between 16-18mm;
2. These phenotypic differences in body mass and beak length are due to differences in their
genes and hence they may be genetically different from one another;
3. This could be due to geographical (different islands) or physiological isolation (beaks
adapted to different food sources) hence leading to reproductive isolation such that they no
longer can breed and form viable offspring, leading to speciation;

(b) Discuss how the information in Fig. 9.1 and Fig. 9.2 shows how these two processes may have
contributed to the evolution of Darwin’s finches.

micro-evolution
1. Microevolution occurred over a few generations after the ancestor population arrived and
adapted to the different food sources with natural selection* selecting for different beak
lengths resulting in changes to the allele frequencies*;
2. This happened on a small scale and no new species are formed;

macro-evolution
1. Macroevolution occurred after many generations spanning a longer period of time where 14
new species are formed due to adaptive radiation;
2. This happened on a large scale where many of the finch populations were geographically
isolated due to rising sea levels, resulting in the disruption of the gene flow* between
populations;
3. Isolated populations evolve independently and with genetic drift and accumulation of
mutations, evolved into new species;
[4]

(c) Fig. 9.3 shows the phylogeny based on whole genome sequencing of the six closely related species
of Darwin’s finches that have evolved most recently.

Describe the advantages of using genome sequences in reconstructing phylogenetic relationships.

[3]
1. Nucleotide data are objective. Molecular character states are unambiguous as A, C, G and T
are easily recognisable and cannot be confused;
2. Nucleotide data are quantitative. Molecular data are easily converted to numerical form and
hence are amenable to mathematical and statistical analysis and hence computation. Degree
of relatedness can be inferred and quantified by calculating nucleotide differences between
species;
3. Nucleotide data can be used to compare species which are morphologically indistinguishable
especially if they are very closely related like the finches;
4. As changes in nucleotide sequences accumulate over time with clockwork regularity. We can
estimate the time of speciation of modern to ancient species and place the speciation events
on a timescale above;

[Total: 10]
10
(a) Describe and explain the changes in the rate of TB transmission that are shown in Fig. 10.1. [3]

1. Trend: As percentage TB vaccination increased drastically from 40% to 100% 1980 to 1989,
the rate of TB transmission per 100 000 decreased sharply from 8.0 to less than 3.7.
2. Trend: From 1989 to 2010, when percentage TB vaccination remained relatively high above
83%, transmission remained relatively low below 5.0 per 100 000;
OR
3. Trend: However the inconsistent percentage of TB vaccination from 1989 onwards resulted in
a base/minimum rate of transmission of at least 3.4 per 100 000
4. Explanation: as a larger percentage of the children are vaccinated, more children were immune
and protected against TB and did not contribute to the transmission of the disease.
5. Quote any complete set of data from both axes

Teacher’s comment:
Many students did not use descriptive words for the shape of the graphs such as sharp or
drastic decrease and steep increase
Students should observe the trend for both graphs is from 1980 to about 1989 and from 1989
to 2010 as the shape of the graph decrease/increase sharply then fluctuates around a certain
value. With only 3 marks should not describe every part of the graph. They should also ignore
the peak at 1987.
When reading from a graph with 2 axes from both sides, students need to be more careful.
Some students read the axis wrongly in % rather than rate or the rate value wrongly e.g 350000
instead of 3.5 per 100000.
Many students mistaken the graph represents herd immunity. For herd immunity, the
transmission rate is much lower. They should take the cue from Fig. 3.2 that Q shows herd
immunity thus Fig. 3.1 where the vaccination fluctuates and goes below 95% with higher rate
of transmission does not.

(b) Suggest reasons for the differences in the rate of TB transmission between country P and country
Q, as shown in Fig. 10.1 and Fig. 10.2. [2]
Either 1 or 2 for 1 mark
1. Country P had a rate of transmission that is erratic and varied between 3.3 and 5.0 per 100 000
from 1989 while country Q has a consistent drop in rate of transmission from 4.5 to 2.5 per 100
000 from 1989;
2. Country Q had a more consistent/robust/compulsory vaccination programme that ensured that
98% of the children were immunised from 1989 while country P could not maintain a high
percentage that at times fell to 83%; - reason 1

Either 3 or 4 for 1 mark

3. This meant that country Q experienced herd immunity*; - reason 2
4. With a sufficiently large proportion of immunised individuals (of above 95%) who cannot spread
the disease, the entire community including the individuals who are not vaccinated/cannot be
vaccinated are protected because transmission is prevented; - explanation of herd immunity
note: the unvaccinated are still vulnerable
 Teacher’s comment:
As this is a comparison question, students must state data clearly from both countries Q and
P. A few students mentioned wrongly comparing Q vs Q instead of Q vs P.
As herd immunity is not established in P, Q should not refer to as showing stronger herd
immunity.
In explanation for pt 4 most students are not able to explain clearly.

[Total: 5]

11
(a) Describe how the concentrations of dengue virus antigen and antibody change during the first ten
days following infection, as shown in Fig. 11.1. [2]
1. As the dengue virus antigen concentration increases steeply from Day 1 to peak at Day 3,
the antibody concentration gradually increases;
2. As the antibody concentration increases steeply/exponentially from Day 3 to plateaus at
Day 10, the virus antigen concentration decreases steadily from Day 3 to zero at Day 9
and 10.

(b) Outline the role of antibodies in eliminating the dengue virus from the blood. [3]
1. Two antigen binding sites per antibody molecule allows agglutination* of the pathogen,
able to bind to two antigens/pathogens at the same time pathogens aggregate/clump to
facilitate clearance
2. Neutralisation* of the pathogen by the antibody which prevents the entry of
pathogen/toxins into the host cells
3. Fc region/constant region of heavy chains bind to Fc receptors on phagocytes to tag /
promote phagocytosis which is opsonisation*;

[Total: 5]
 2018 H2 A level Paper 3
1(a) (i) Describe the patterns shown in Table 1.1. [2]
1. As the degree of differentiation / specialization increases from blood stem cell from bone
marrow to mature B lymphocyte from lymph gland,
2. the degree of DNA methylation decreases from 1.00 to 0.61 arbitrary units(AU),
3. and the degree of gene expression increases from 1.0 to 5.5AU.

(ii) Explain how DNA methylation changes gene expression. [3]

1. addition of methyl group to selected cytosine* nucleotides in on DNA (e.g. a CG
sequence);
2. Results in condensation of chromatin fibres.
3. Reduces the assessiblity of promoter, hence prevents the binding of general
transcription factors* and RNA polymerase* to promoter* / preventing formation of
transcription initiation complex*
4. Preventing transcription of the gene;

(iii) Compare the data for the cancerous B lymphocytes in Table 1.2 with the data in Table 1.1
and discuss what this suggests about the nature of the cancerous B lymphocytes. [4]
1. Both the degree of DNA methylation (0.95AU) and gene expression (1.5 AU) for
cancerous B Lymphocytes are in between these values blood stem cell from bone
marrow and immature B lymphocyte from bone marrow.
2. High levels of DNA methylation, which results in high levels of condensation of chromatin
in the genome.
3. Low level of genes expression, only 1.5 AU, hence likely only housekeeping genes are
expressed / gene that codes to proteins that are required on all cells, e.g. ribosomal
genes.
4. Hence cancerous B lymphocytes is expected to largely undifferentiated, similar to that of
the blood stem cell.
5. And that many tissue-specific genes and proteins, e.g antibodies are not likely to
expressed.

(b) (i) Describe and explain the potential consequences of the cell and individual person of
methylation of the promoter region of the p53 tumour suppressor gene. [4]
1. Methylation at promoter region of p53 tumour suppressor gene results in condensation
of chromatin in the promoter region.
2. Hence general transcription factors(GTF) and RNA polymerase(pol) cannot bind to the
promoter to form the transcription initation complex, hence cannot initiate transcription
of the p53 gene / p53 gene note expressed.
3. p53 protein cannot function as a specific transcription factor and cannot activate genes
that are involved in DNA repair, cell cycle arrest and stimulating damaged cell to undergo
apoptosis;
4. Cell cycle continues without repairing DNA/ cell does not undergo apoptosis;
5. Results in uncontrolled cell division / tumour formation
 (ii) 5-azacytidine is a chemical that inhibits the enzyme DNA methyltransferase. This enzyme
adds methyl groups to DNA.

Explain, with reference to Fig. 1.2, why 5 azacytidine may be useful in treating cancers in
older people. [3]
5 azacytidine inhibits enzyme DNA methyltransferase and
1. hence preventing methylation at promoter region of p53 tumour suppressor gene,
preventing condensation of chromatin in the promoter region.
2. Hence allowing GTF and RNA pol to bind to p53 promoter / form the transcription
initiation complex at promoter, hence initiating transcription of the p53 gene /
expression of p53 gene.
3. p53 protein produced function as specific transcription factor and cannot activate
genes that are involved in DNA repair, cell cycle arrest and stimulating damaged cell
to undergo apoptosis;
4. Preventing uncontrolled cell division treating cancer.

(c) (i) Telomere DNA is tightly condensed due to histone modification. DNA methylation would
have a similar effect on the packaging of the DNA, but DNA methylation is not possible at
telomeres.

Explain why telomere DNA cannot be methylated. [1]

1. Telomeres do not have the nucleotide sequences recognized by DNA

methyltransferase for DNA methylation / Lack of methylation site
R: lack of cytosine residues!!!

(ii) Outline two functions of a telomere containing hundreds of repeat sequences. [2]
1. Telomeres are non-coding* tandem repeat sequences found at both ends of linear
chromosomes;
(non-coding must be mentioned somewhere in answer)

Role – main
2. Each round of DNA replication will result in the shortening of daughter molecules at the
telomeres because DNA polymerase is unable to replace the RNA primers with DNA;
(idea of end replication problem)
3. Since telomeres are non-coding, this ensures that vital genetic information/genes are
not lost / eroded with each round of replication; wtte

Role – others
4. By forming a loop with 3’ overhang, they protect and stabilise terminal ends of
chromosome, hence preventing fusion of the ends with those of other chromosomes;
5. Either: prevent triggering pathways that lead to cell arrest or cell death, because
exposed 3’ overhang will be perceived as DNA damage/DNA double strand break;
OR:
prevent DNA repair machinery from recognising the ends of chromosomes as DNA
breaks/damage, hence preventing apoptosis;
6. Either: The 3’overhang of the telomeres allow their own extension, by providing an
attachment point for the correct positioning of the enzyme telomerase in certain cells,
e.g. germ cells
 OR:
They possess a 3’ overhang which base pairs with the RNA template on telomerase, so
ensures proper alignment of telomerase and allows extension of telomeric ends in
certain cells e.g. germ cells.

(iii) Most human cell lines stop dividing after 40 to 60 rounds of cell division. This is known as
the ‘Hayflick limit’ and is due to changes in the DNA in telomeres.

Cancer cell lines and stem cell lines, in contrast, can divide indefinitely without limit.

Explain how changes in the DNA in telomeres prevent most human cell lines from dividing
beyond the Hayflick limit and suggest how cancer cells and stem cells are able to overcome
this limit. [4]
1. Each round of DNA replication will result in the shortening of daughter molecules at the
telomeres at the 5’end
2. because DNA polymerase is unable to replace the RNA primers with DNA nucleotides
3. In stem cells and cancer cells, expression of telomerase* gene / Presence of
telomerase*
4. which extends telomere / hence telomere length to be maintained / prevents telomeres
from reaching critical length / Hayflick limit / thus; wtte
5. Allowing them to undergo continuous cell division to allow many replication cycles to
occur / prevents apoptosis;

(d) (i) With reference to Fig. 1.4, evaluate the extent to which telomere length can predict a
person’s chronological age. [3]

1. In general, there is a negative relationship between chronological age and average

telomere length / the as the chronological age increases from 18 to 78, the average
telomere length decreases linearly from 7.8kb to 6.2kb.
2. However, there are wide spread / deviation of telomere length for each age group,
3. with length of telomere for some individuals deviating significantly from that of their
average age group,
e.g. 31 yo individual with 6.3kb telomere length, which is the average length for
people of age 78.
E.g Individuals of age 60 having a telomere length of 8.5kb, which is significantly
higher than averge 20 yo individual.
4. Hence may not be a reliable prediction.

(ii) Suggest what this implies for the health and life expectancy of person P. [4]
1. The DNA methylation age of Person P of ard 87years, is significantly higher than of the
average of 58 years for his age of 58 years.
2. This suggests that significantly high amount of his cells being undifferentiated,
3. Hence an indication that higher chance / tendency of the promoter region of the p53
tumour suppressor gene being methylated, hence p53 not expressed.
4. Suggesting a high chance / tendency of person P suffering from cancer thus may have
a lower life expectancy than the average 58 year old.

[Total: 30]
2(a) Outline the components of the non-specific immune system in mammals. [4]
1. Impermeable anatomical barriers such as intact skin and mucous membrane that prevents
pathogens from entering the organism;
2. Chemical barriers that include secretions in that has antimicrobial substances such as lysozyme
that cleaves glycosidic bonds of peptidoglycan cell walls of bacteria, or acidic pH that denature
proteins in pathogens;
3. Phagocytes such as neutrophils, macrophages and dendritic cells that make up the cellular
component of the non-specific immune system that engulf pathogens by phagocytosis;
4. Macrophages are also responsible for the inflammatory response as they secrete chemokines
to recruit neutrophils to the site of infection to carry out phagocytosis;
5. Macrophages secrete cytokines that increases permeability of blood vessels allowing
neutrophils to migrate into tissue from the blood;

(b) Complete Table 2.1 with ticks to show whether these two example of enhanced immunity in
offspring are active, natural or passive or a combination of these.

Table 2.1

type of immunity mammal honey bee

active
natural
passive
Active in honey bee because the immune response of the recipient is stimulated and enhanced
by exposure to pathogen even though no antibodies are produced;
[2]

(c) Honey bees are important for pollinating fruit crops. Without pollination, fruits are not produced.
Suggest why it is economically important to investigate the mechanism of immunity in honey bees.
[3]
1. It is important to protect bees from pathogens, so that pollination of fruit crops could continue
to produce fruits and generate revenue; draw link b/w immune system to economy
2. An understanding of the bee’s mechanism of immunity will allow us to find ways to manipulate
or enhance the bee’s immune system conferring long term protection against pathogens;
3. E.g. Vaccinate the bees against pathogens by exposing the eggs to fragments of the pathogen;
4. E.g. Stimulate the cells of the non-specific immune system to respond strongly to pathogens;
similar to pt 3
5. E.g. Stimulate proliferation of cells in the non-specific immune system to combat infectious
agents;

1 mark for an example that makes sense. Pt 1 and 2 compulsory for 3marks.
[Total: 9]

3(a) Explain why environmentalists would like all new coal-fired power stations to include CCS or CCU
facilities. [4]
1. CO2 is a greenhouse gas that will trap infra-red radiation from escaping from the atmosphere
that results in global warming;
2. Thus environmentalists would like CCS and CCU to be used to limit CO2 emission so as to
mitigate the effect of global warming:;
3. to prevent coral bleaching as a result of high ocean temperatures;
4. to prevent more extreme weather conditions such as drought in some areas and flood in
others as a result of heavy rainfall;
(b) Evaluate the effectiveness of the processes shown in Fig. 3.1 as a solution to the problem of carbon
dioxide emissions from burning fossil fuels. [3]
1. CCU uses CO2 from coal burning to extract oil from oil field thus reduces CO2 emission from
burning fossil fuels to provide energy for the process;
2. With CCS CO2 is stored underground rather than release to the atmosphere thus reducing the
emission of greenhouse gases into the atmosphere;
3. However the processes of carbon dioxide transport and pumping may incur extra energy use
which results in CO2 emission from fossil fuels;
4. Furthermore the oil extracted will be used for energy requiring processes such as transport
which will add CO2 to the atmosphere;

(c) The release of carbon dioxide into the atmosphere from coal-fired power stations must be
considered in relation to other processes that affect the concentration of carbon dioxide in the
atmosphere.

For example, respiration also adds carbon dioxide to the atmosphere.

Outline how carbon dioxide is produced in respiration. [4]

1. Carbon dioxide is produced in the link reaction* and Krebs cycle* of aerobic respiration in the
mitochondrion*;
2. During link reaction 3C pyruvate* undergoes oxidative decarboxylation and the addition of
coenzyme A to produce 2C acetyl coA* and carbon dioxide;
3. In the Krebs cycle and 6C citrate* undergoes oxidative decarboxylation to produce 5C α-
ketoglutarate* and is further decarboxylated to form 4C oxaloacetate* with the release of
carbon dioxide;
4. Also during alcohol fermentation in yeast, 3C pyruvate* is decarboxylated /carbon dioxide is
released to form 2C ethanal*;

[Total: 9]

4(a) Describe the process by which a C3 plant makes sugars in its leaves using sunlight, water and
carbon dioxide. [15]
Light dependent reaction
1. Light energy* from the sun is absorbed by accessory pigments molecules in light harvesting
complex of photosystems (PS) I and II*;
2. The electrons in these pigment molecules become excited and, when returning to their ground
states, pass on the released energy to the next pigment molecule and excite the electrons
present in them, as a result. This resonance transfer of energy occurs until it reaches one of
the two special chlorophyll a molecules (P700 in PS I & P680 in PS II) in the reaction centre;
3. When the special chlorophyll a molecule absorbs the energy, an excited electron is displaced
(Chl a chl a+ +e-) leaving an electron hole in PS I and II. This electron is captured by a
primary electron acceptor molecule in the reaction centre;
4. The electron hole in PS II is replaced with electrons from the photolysis of water to give H+
ions, electrons and O2.
5. As the excited electrons flow down a chain of carriers of ETC with progressively lower energy
levels
6. energy lost is coupled to pumping H+ ions across the membrane into the thylakoid space.
7. H+ accumulates in the thylakoid space which serves as the H+ reservoir. In this way, the electron
transport chain transforms redox energy to a proton-motive force*.
8. As protons / H+ diffuse down the concentration gradient back into the stroma via the ATP
synthase complex*.
 9. ADP is phosphorylated to ATP* in the process via chemiosmosis*.
10. In non-cyclic photophosphorylation, the final electron acceptor* is NADP* which is a
coenzyme.
11. Reduced NADP (NADPH) carries the electrons which are used in the Calvin cycle in the stroma
of the choloroplast.

Light independent reaction/Calvin cycle

Carbon fixation:
12. During carbon fixation stage, CO2 is combined with ribulose bisphosphate (RuBP)* to form
an unstable 6 carbon molecule which;
13. splits up immediately into 2 molecules of glycerate phosphate (GP)*;
14. The enzyme catalysing this reaction is RuBP carboxylase (RUBISCO)*;

Reduction by NADPH:
15. NADPH (from light-dependent reaction) is the reducing power used to reduce* GP to
glyceraldehyde-3-phosphate (G3P);
16. ATP (from light-dependent reaction) is the source of energy required;
17. G3P is the first sugar formed in photosynthesis and the end product of Calvin cycle;

Regeneration of RuBP:
18. 5 molecules of G3P are used to regenerate* 3 RuBP so that the cycle of carbon dioxide fixation
can continue. This requires 3 ATP (from light-dependent reaction);
19. The net synthesis of 1 molecule of G3P requires 3 CO2 to be fixed;
20. 2 molecules of G3P may be used to form 1 molecule of glucose (hexose sugar);

(b) Germination is the process by which a seed with access to water, oxygen and a suitable
temperature begins to grow into a young plant. A seed consists of an embryo and a food store.

The mass of a soaked seed undergoing germination in the light was recorded. In the first five days,
the mass of the seed decreased. In the next five days as the young plant grew, this mass was
regained.

Explain the decrease and gain in mass as the seed germinated and grew into a young plant over
this period of ten days. [10]
1. The food store present in most seeds is in the form of carbohydrates, fats and proteins;
2. The stored food is used to support the embryo during seed germination;

First 5 days of seed germination:

3. Imbibition of water activates hydrolytic enzymes* allowing metabolic activity to resume in the
rehydrated seed;
4. The hydrolytic enzymes break down the proteins into amino acids, fats into lipids and the
starch into glucose;
5. The amino acids and lipids are transported to the embryo where it is used during cell division
and growth of the embryo;
6. Seed begins to undergo aerobic respiration* as oxygen enters and the glucose is broken
down into energy in the form of ATP, carbon dioxide and water;
7. As the embryo is still developing, photosynthesis does not occur and the mass is lost mainly
in the form of carbon dioxide during the link reaction and Krebs cycle of aerobic respiration;
 From day 6 to day 10 of seed germination:
8. As the embryo develops into a young plant, leaf development and chlorophyll synthesis takes
place;
9. The young plant is able to undergo photosynthesis* during which glucose is synthesised;
10. The glucose produced is converted to starch where it is stored in different parts of the plant;
11. The glucose produced is also used in cellulose synthesis which is the main component of plant
cell wall in newly formed cells;
12. The synthesis of glucose, starch and cellulose in the developing plant will cause an increase
in the overall mass of the plant;

[Total: 25]

5 (a) Birds are thought to have descended from dinosaurs.

Explain how a population of a species of dinosaur could change over time into a new species with
bird-like features. [15]

1. There exist variation* in a population of dinosaurs for natural selection* to act on;
2. where some have more distinct bird-like features like wings than others;
3. If the environment changes, such that there is a new selection pressure*;
4. such as new predator in the area;
5. those with favourable traits such as wings that enable flight which lets them escape from the
predator;
6. will have a selective advantage to the local conditions and will be selected for,
7. and will survive, reproduce and pass on their alleles to the next generation;
8. increasing frequency of favourable alleles in the population;
9. and hence microevolution* would have occurred;
10. If this population is geographically isolated from other populations of the same species where
there are no predators/ different selection pressures/niches;
11. disruption of gene flow* will occur between the two sub-populations;
12. The two sub populations will evolved independently of each other;
13. their allele frequencies will change as they accumulate different genetic mutations, and are
subjected to genetic drift and natural selection;
14. Over a hundreds and thousands of generations, each population will become reproductively
isolated*
15. Such that they can no longer interbreed* to produce viable, fertile* offspring;
16. and hence new species are formed through allopatric speciation and
17. hence macro evolution would have occurred;
(b) Dinosaurs are now extinct but there are approximately 10 000 species of living birds. It is thought
that birds evolved from small dinosaurs and that being small helped their survival during a time of
rapid environmental change when species of large dinosaur went extinct.

Suggest and explain why, in a rapidly changing environment, small animals may be at an
evolutionary advantage compared to large animals. [10]
1. A) Small animals populations put only a small investment of resources into each offspring;
B) so that there will be enough resources to produce more offspring;

2. A)Small animals populations have high reproductive output;

B) so that there is greater chance of survival in a rapidly changing environment;

3. A) Small animals populations are also generally not very invested in protecting or rearing their
young for many years;
B) Often, the eggs are fertilized and then cared for only a short period of time. The benefit of
this strategy is that if resources are limited or unpredictable, you can still produce some young;

4. A) Small animal populations grow rapidly, with shorter lifespans;

B) this short generation time allow the fittest to survive, reproduce and pass on their favourable
alleles to the next generation, allowing faster rate of evolution to adapt to the changing
environment;
5. A)The young of small animals tend to be more rapidly maturing and develop early
independence;
B) This allows less dependence on parents so even if parents do not survive the changing
environment, there is a chance for their offspring to adapt and survive;
6. A) Small animal populations require less food as compared to large animals or maybe able to
avoid predation due to their size;
B) hence allow them a selective advantage over the larger animal populations;

[Total: 25]
2019 H2 A level Paper 2

Section A
Answer all the questions in this section.

1 Bacteria divide by binary fission. The time taken to divide into two cells is called the generation
time.

Fig. 1.1 is a diagram of a dividing cell of the bacterium Escherichia coli.

Fig. 1.1
(a) List three structural features of E. coli that are visible in Fig. 1.1 [3]

1. Ribosomes
2. Circular DNA
3. (Peptidoglycan) Cell wall
4. Cell surface membrane

(b) For one of the structural features listed in (a), state a detail of this feature that is
characteristic of a bacterial cell. [1]
1. Ribosomes – 70s ribosomes in bacteria

2. DNA - circular DNA that is double stranded

OR
DNA molecule is not associated with histone but other histone-like protein
molecules.

3. Cell wall – Made up of peptidoglycan* polymers

4. Cell surface membrane - Phospholipid bilayer with electron transport chains and
ATP synthase are embedded to produce ATP

(c) Describe what happens to the genome of bacteria during binary fission. [2]
1. Bacterial genome which is made of DNA replicates by semi-conservative
replication*
2. DNA is unzipped by breaking hydrogen bonds between bases of the 2 strands
3. each parental strand serves as a template for synthesis of daughter strands
4. free deoxyribonucleotides complementary base pair with bases on the parental
/template strand
5. DNA polymerase* forms phosphodiester bonds* between adjacent
deoxyribonucleotides

In a study, E. coli bacteria were cultured under a variety of nutrient conditions. After several
generations, the generation time and the mean mass of an individual bacterial cell in each
culture were measured. Table 1.1 shows the results of this study.

Table 1.1

2019 H2 Biology A Level Paper 3

[Turn over
 2

mean mass of
nutrient generation time
individual bacterial
availability /minutes cell /10-15 g

low 100 150

60 260
40 430
30 640

high 24 870

(d) Describe how the generation time and mean mass of an individual bacterial cell changes
with nutrient availability, as shown in Table 1.1. [2]
1. As nutrient availability increased from low to high, the generation time decreased
from 100 to 24 minutes (quote data);
2. And mean mass of individual bacterial cells increased from 150 x 10-15 g to 870 x 10-
15
g (quote data).

(e) Suggest an explanation for the effect of decreasing nutrient availability on the generation
time of these bacteria, as shown in Table 1.1. [2]
Name nutrient + Effect of a lack of these nutrients on named cell processes

1. With low or no glucose, the bacteria will be unable to carry out aerobic respiration to
generate sufficient levels of ATP
2. Hence the bacterial cells will have insufficient energy required for cell division/binary
fission and hence take a longer generation time

OR

1. Certain essential amino acids, which cannot be produced by the bacterial cells, need
to be taken up from the environment
2. Without which the bacterial cells will be unable to synthesize important bacterial
proteins for growth, and hence require a longer generation time

AVP

[Total: 10]

2019 H2 Biology A Level Paper 3

 3

2 Cellulose is a polymer made up of monomers bonded, together.

(a) Name the monomers that join to form a cellulose molecule. [1]
β-glucose monomers

(b) Name the covalent bond between two of these monomers in a cellulose molecule and
describe how this bond is formed. [3]
1. β(1-4) glycosidic bond*
2. The glycosidic bond forms between C1 of one β glucose* monomer and C4 of
another β glucose* monomer;
3. Condensation* reaction joining two β glucose residues with removal of one water*
molecule;
4. Reaction is catalysed by enzymes*;

(c) Explain how the molecular structure of cellulose is related to the high tensile strength of
the cellulose cell walls of plants. [3]
1. Adjacent β glucose* monomers are rotated 180o* with respect to each other
cellulose
2. Forming a straight* chain/molecule and is able to lie parallel* to other cellulose
chains;
3. Hydroxyl groups* project outwards from each molecule, allowing extensive
hydrogen bonding* between adjacent chains, forming microfibrils* with high
tensile strength* that make up the cell wall

2019 H2 Biology A level Paper 3 [Turn over

 4

Fig. 2.1 shows the enzyme cellulose synthase complex in a plant cell surface membrane. The
enzyme complex synthesises the eighteen cellulose molecules that form a microfibril of the
cellulose cell wall.

Fig. 2.1

(d) Suggest why cellulose must be synthesised at the cell surface membrane and not inside
the cell. [3]
1. Cellulose is a macromolecule found outside the cell as part of the cell wall and
therefore easier to simply deposit it there.
2. Cellulose molecule may be too large to pass through cell membrane if it has to be
transported to the exterior of the cell.
3. Cellulose is insoluble in the hydrophobic core of the phospholipid, and is hence
synthesized at the cell surface membrane and transported out of the cell

[Total:10]

2019 H2 Biology A Level Paper 3

 5

3 Fig. 3.1 is an electronmicrograph showing simultaneous transcription and translation of a single

gene in the bacterium Escherichia coli. The arrow shows the place where transcription of the
gene begins.

Fig. 3.1

(a) Explain why transcription and translation are able to occur simultaneously in a bacterial
cell such as E. coli. [2]
1. E.coli is a prokaryotic cell that does not contain a membrane bound nucleus/nuclear
envelope. Hence as soon as the mRNA is transcribed the ribosomes can attach to
the mRNA and translation can be carried out;

2. Also, unlike eukaryotic cells, DNA that is transcribed to mRNA in prokaryotic cells
need not undergo post-transcriptional modification and hence transcription and
translation can occur simultaneously;

Fig. 3.2 is a diagram showing events just to the right of the arrow in Fig. 3.1.

Fig. 3.2

(b) Identify structures A, B and C, as shown in Fig. 3.2. [3]

A: DNA double helix
B: mRNA/messenger RNA
C: polypeptide

2019 H2 Biology A level Paper 3 [Turn over

 6

(c) Describe how structure D is different from structure E. [2]

1. Structure D is made up of rRNA and proteins, while structure E is made of proteins.
2. Structure D is comprised of 2 (ribosomal) subunits, while structure E is comprised of
1 subunit
3. Structure D uses mRNA as a template to synthesize polypeptides, while structure E
uses DNA as a template to synthesize mRNA.

(d) List three ways in which the process of transcription is different from the process of
translation. [3]
Transcription Translation
Location Occurs in nucleus* of Occurs on ribosomes* in
eukaryotes cytoplasm / rough
endoplasmic reticulum

Process DNA used as template* in mRNA used as template*

synthesis of mRNA in synthesis of polypeptide

Enzymes that joins Enzyme RNA Peptidyl transferase* on

monomers polymerase* links ribosome joins amino
ribonucleotides acids

Linkages Ribonucleotides linked by Amino acids linked by

phosphodiester bonds* peptide bonds* to
to form mRNA produce polypeptide

Product Single-stranded mRNA Polypeptide

Direction read DNA template read from a mRNA read from 5’ to 3’

3’ to 5’ direction direction

[Total: 10]

2019 H2 Biology A Level Paper 3

 7

4 Fig. 4.1 shows a transmission eIectronmicrograph of viruses similar to the T4 phage infecting
a bacterial cell.

Fig. 4.1

(a) Identify structures X, Y and Z on Fig. 4.1. [3]

X: (Icosahedral) Capsid head
Y: (contractile) Tail sheath
Z: Tail fibre

(b) With reference to Fig. 4.1, outline how the T4 phage viral genome is replicated. [3]
1. Inside the cell, the T4 bacteriophage DNA is immediately transcribed to synthesise
messenger RNA using the host RNA polymerase
2. Phage enzymes coded by the phage genome takes over the bacterium’s
macromolecular (protein, RNA, DNA) synthesising machinery for its own use.
3. The phage uses the host cell’s nucleotides and DNA polymerase to synthesise many
copies of phage DNA.

Table 4.1 shows a comparison of the energy requirements of the reproductive cycle of a T4
phage and the energy requirements of the reproductive cycle of an influenza virus.

Table 4.1

2019 H2 Biology A level Paper 3 [Turn over

 8

(c) Describe the two main differences between the energy requirements of the reproductive
cycle of a T4 phage and the energy requirements of the reproductive cycle of an influenza
virus, as shown in Table 4.1.

Suggest an explanation for each of the two differences. [4]

difference 1 and explanation:

1. T4 phage requires much higher energy, of 200 a.u., than influenza virus, of 10 a.u.
for genome replication
2. T4 could require more energy due to the larger size of the viral genome required to
be replicated

OR
T4 viral DNA could require more energy to replicate due it being double stranded

OR
T4 requires more energy to degrade host cell DNA to synthesize viral DNA as
opposed to influenza virus which synthesizes viral mRNA in animal host cell using
existing ribonucleotides

difference 2 and explanation:

1. T4 phage requires lesser/no energy/0 a.u for viral exit, as compared to influenza
virus, requiring 300 a.u. for viral exit.
2. T4 viral gene codes for lysozyme which causes host bacterial cell wall to lyse and
release bacteriophages, thus not requiring energy. However the influenza virus will
need to bud off from host cell and acquire host membrane, thus requiring more
energy.

[Total: 10]

2019 H2 Biology A Level Paper 3

 9

5 Table 5.1 shows the gene mutation that leads to sickle cell anaemia and some of the
consequences of this gene mutation.

Table 5.1

(a) Describe the gene mutation shown in row 1. [2]

1. A single base substitution*
2. From thymine to adenine

(b) Describe how the gene mutation shown in row 1 leads to the change in row 3. [3]
1. The change in the nucleotide / base sequence in DNA* from thymine to adenine
2. Results in a change in sequence of the mRNA, and a change in codon*
3. Change from GAG coding for amino acid glutamate to GUG which codes for valine

(c) Explain how the change in row 3 affects the haemoglobin molecule. [3]
1. Hydrophilic* charged glutamic acid* is replaced by hydrophobic*, non-polar
valine*;
2. This changes the primary, secondary and tertiary structure* because the way
polypeptide chain folds is affected by change in R groups* and bonds formed;
3. At low O2 concentrations, loss of O2 from HbS* results in an unusual conformational
change that causes a hydrophobic patch to stick out;
4. This hydrophobic patch attaches to a hydrophobic patch on another HbS causing
them to crystallise / polymerise into insoluble fibers*;

(d) Explain how this change to haemoglobin results in the symptoms of sickle cell anaemia
when oxygen concentration is low. [2]
1. Long insoluble HbS fibers within red blood cell causes its shape to be distorted
from a normal biconcave shape to a sickle* shape;
2. Sickle red blood cells are more fragile resulting in them having a shorter life span,
results in shortage of red blood cells and poor oxygen transport resulting in
anaemia;
3. Sickle-shaped red blood cells, being pointed and elongated, may also get lodged in
small blood vessels (capillaries) and therefore interfere with blood circulation. This
may result in organ damage;

[Total: 10]

2019 H2 Biology A level Paper 3 [Turn over

 10

6 Western honey bees, Apis mellifera, live in colonies. Colonies consist of three types of bees:

• a single fertile female called a queen bee

• infertile females, called worker bees
• fertile males, called drones.

Queen bees and worker bees develop from fertilised eggs so are diploid. Drones develop from
unfertilised eggs so are haploid.

Within each colony, the single queen bee lays eggs in small compartments called cells. The
eggs hatch into larvae that develop within the cells. When the larvae are fully grown, the cells
are capped by worker bees. The larvae complete their development into adult bees inside these
capped cells.

In artificial hives, larvae are susceptible to a number of diseases that can spread through the
colony. The spread of these diseases can be reduced by two particular behaviours of worker
bees, each controlled by a single gene.

One gene has two alleles, A and a. The recessive allele causes worker bees to detect and
uncap any cell that contains a diseased larva.

The other gene has two alleles, B and b. The recessive allele causes worker bees to remove
larvae from cells that have been uncapped and discard them from the hive.

() A queen bee that is heterozygous for both genes is crossed with a drone that has the
recessive allele for both genes. After fertilisation, the queen bee is able to lay both
fertilised and unfertilised eggs.

Fig. 6.1 shows all possible male and female offspring of this queen bee.

Fig. 6.1

2019 H2 Biology A Level Paper 3

 11

Draw genetic diagrams to explain the results shown in Fig. 6.1 for the:

(i) Male offspring

Parental phenotype: Fertile female
Parental genotype: AaBb

Gametes: AB Ab aB ab

Offspring genotype: AB, Ab, aB, ab

Offspring phenotype: All fertile males (drone)
[2]
(ii) Female offspring
Parental phenotype: Fertile female (queen) X Fertile male (drone)
Parental genotype: AaBb X ab

AB Ab aB ab ab
Gametes:

Offspring genotype: AaBb Aabb aaBb aabb

Offspring phenotype: All female

[3]

(b) It is possible to selectively breed western honey bees so that all worker bees in a hive
display the ’uncapping cells’ behaviour and the ‘removing larvae’ behaviour.

Circle two of the offspring on Fig. 6.1 to identify which bees should be used in a cross to
ensure that all worker bees in the next generation show both of these behaviours to
reduce disease.

See Fig. 6.1 – Thinking: One fertile queen bee, one fertile drone. Both must result in
homozygous recessive offspring.
[2]
(c) Explain how the environment determines whether fertilised eggs develop into queen
bees or worker bees. [3]

1. Diet of the larvae determines if it develops into queen bee or worker bee
2. Larvae fed royal jelly*, a protein-rich secretion from young worker bee will develop
into a queen bee.
3. Chemicals in royal jelly trigger expression of genes involved in the development of
ovaries and reproductive organs.
4. Larvae fed a nectar and pollen will develop into worker bee.

[Total: 10]

2019 H2 Biology A level Paper 3 [Turn over

 12

7 (a) Describe the role of oxygen in aerobic respiration. [3]

1. Oxygen is final electron acceptor* at the end of electron transport chain*, where
it will combine with electrons and protons to form water (accept: 2e─ + 2H+ + ½O2
H2O);
2. By removing electrons, oxygen re-oxidises electron transport chain so that NADH*
and FADH2* can continue to donate electrons to the chain, thereby allowing oxidative
phosphorylation* to continue to produce ATP*;
3. This allows regeneration* of NAD+* and FAD* allowing them to pick up more
electrons and protons from glycolysis*, link reaction* and Krebs cycle* to keep
them going;

(b) In yeast cells, less glucose is required to maintain cell metabolism under aerobic
conditions than under anaerobic conditions.

Explain why. [3]

1. Under aerobic conditions, complete breakdown of glucose produces 38 ATP*
molecules per glucose molecule as compared to 2 ATP* molecules in anaerobic
respiration;
OR
under anaerobic conditions, 19 glucose molecules will be needed to generate same
amount of energy that 1 glucose molecule can yield under aerobic conditions;

2. Under aerobic conditions, link reaction* and Krebs cycle* produce NADH* that will
be oxidised and hence regenerated by the electron transport chain* during
oxidative phosphorylation*, generating additional ATP;

3. However, under anaerobic conditions, glycolysis only produces a net of 2 ATP

molecules* for each glucose molecule oxidized and NAD+* and is only regenerated
through fermentation processes to allow only glycolysis* to continue.

2019 H2 Biology A Level Paper 3

 13

PFK (phosphofructokinase) is an enzyme that catalyses one of the steps in glycolysis. Its
substrate is fructose 6-phosphate. ”

Fig. 7.1 shows the effect of increasing substrate concentration on PFK activity at high and low
concentrations of ATP.

Fig. 7.1
(c) With reference to Fig. 7.1, describe the effect of a high concentration of ATP on PFK
activity. [2]

1. High levels of ATP slows down / inhibits PFK activity (as the graph is below the graph
represented by low ATP);
2. This inhibition is most pronounced at lower substrate concentrations + Quote value
from graph;
3. this inhibition is overcome by high substrate concentrations + Quote value from
graph;
(NB: despite the observation in points 2 & 3, do note that ATP is NOT a competitive
inhibitor, since PFK is an allosteric enzyme - meaning that ATP would be binding to an
allosteric site)

2019 H2 Biology A level Paper 3 [Turn over

 14

(d) Using the information in Fig. 7.1 and your subject knowledge, suggest a mechanism to
explain how ATP controls the rate of glycolysis. [3]

1. (low levels of ATP promote PFK activity); When ATP* levels are high, it will bind to
allosteric site of PFK;
2. This alters 3-D conformation* of enzyme’s active site*, such that its substrate is no
longer complementary in shape and charge* to active site, and this stabilizes the
inactive conformation of PFK which will have a lower affinity for substrate;
3. resulting in a decrease in rate of glycolysis due to end-product inhibition;

[Total: 11]

2019 H2 Biology A Level Paper 3

 15

8 Recently a form of insulin has been developed 1hat can be administered orally instead of by
injection.

Fig. 8.1 shows how the blood insulin concentration of diabetic rats changes over time after
equivalent doses of injected insulin and oral insulin are administered.

Fig. 8.1

Fig. 8.2 shows how the blood glucose concentration of diabetic rats changes over time after
equivalent doses of injected insulin and oral insulin are administered.

Fig. 8.2

2019 H2 Biology A level Paper 3 [Turn over

 16

(a) Describe how the blood insulin concentration changes after administering insulin orally,
as shown in Fig. 8.1. [2]

1. From 0 minute to 9 hours, the blood insulin concentration increase gradually from 4
milliunits dm-3 to 64 milliunits dm-3
2. From 9 hours to 25 hours, the blood insulin concentration decrease gradually from
64 milliunits dm-3 to 12 milliunits dm-3

(b) Explain how Fig. 8.1 and Fig. 8.2 provide evidence that insulin controls blood glucose
concentration. [2]

1. Ref to inverse relationship between concentration of insulin and blood glucose

concentration
2. After oral administration of insulin, the insulin concentration increases from from 4
milliunits dm-3 at 0 hours to 64 milliunits dm-3 at 9 hours and this results in a decrease
in blood glucose concentration from 400 mg dm-3 at 0 hours to 100 mg dm-3 at 9
hours
3. When insulin concentration decreases from from 64 milliunits dm-3 to 12 milliunits dm-
3
and this results in a increase in blood glucose concentration from 100 mg dm-3 at 9
hours to 320 mg dm-3 at 7 hours

Accept if student quota data from injected insulin.

(c) Suggest two advantages of using orally administered insulin in treating diabetes. [2]

1. Does not require injection of insulin, thus lessen pain for patients / decreases the risk
of infections due to injections.
2. Has longer lasting effects thus keeping the blood glucose level lower for a longer
period of time.

Fig. 8.3 shows the molecular structure of the insulin tyrosine kinase receptor on the surface of
a target cell, without insulin attached (left-hand diagram) and with insulin attached (right-hand
diagram).

2019 H2 Biology A Level Paper 3

 17

Fig. 8.3

(d) Explain how the binding of insulin to the insulin tyrosine kinase receptor triggers a
response in the target cell that reduces the blood glucose concentration. [3]

1. Conformational change in the intracellular domain of receptor results in activation

of intrinsic tyrosine kinase;
2. Intrinsic tyrosine kinase activity of each subunit in the intracellular domain cross-
phosphorylates /autophosphorylates the tyrosine* residues on the other subunit;
3. Other relay proteins inside the target cell will be able to bind to the phosphorylated
tyrosine residues and become activated themselves;
4. This will enable the signal to be transduced within the cell until an appropriate
cellular response is reached.

Cellular responses include :

5. Increase the number of glucose transporters (GLUT4) on the plasma membrane;
6. increasing the permeability of the plasma membrane to glucose to increase
uptake of glucose from the blood by these cells, causing blood glucose
concentration to drop;
7. increased activation of the enzyme glycogen synthase, which catalyses the
synthesis of glycogen

2019 H2 Biology A level Paper 3 [Turn over

 18

9 Ecomorph is a term first used by Ernest Edward Williams in 1972 to describe those ‘species
with the same structural habitat/niche’ that are ‘similar in morphology and behaviour, but not
necessarily close phyletically’ (phylogenetically).

Eleven closely related species of insect-feeding Anolis lizards occur across three of the largest
islands in the Caribbean: Puerto Rico, Cuba and Jamaica. Each island has its own particular
mix of species and none of these species occur on more than one of the islands.

During the evolution of these eleven species, each evolved into one of four different ecomorphs:
canopy, twig, trunk-ground or grass-bush. Each ecomorph occurs in a particular habitat and
has specific adaptations.

The four ecomorphs, and their main adaptations, are represented in Fig. 9.1. The four
ecomorphs are drawn to the same scale.

Fig. 9.1

(a) Describe briefly why these four ecomorphs have such different adaptations despite all
the species of Anolis lizards in these ecomorphs being closely related. [3]
1. Random mutations resulting in new alleles as well as sexual reproduction, meant
that there was variation* in size, leg length and tail length in the population;
2. Selection pressure*, possibly the availability of insects (food) in different sections of
the tree or a need for escaping ground-based predators resulted in selection of
different leg lengths or toe pads; (Give at least one context specific explanation)
3. These various ectomorphs of lizards survived and reproduced and passed on their
alleles* their offspring; R: genes
4. Over time frequency of allele responsible for the various favourable adaptations
increased as a result

2019 H2 Biology A Level Paper 3

 19

Fig. 9.2 shows the phylogeny based on DNA sequence data for the eleven species of
Anolis lizards that comprise these four Ecomorphs.

Fig. 9.2

(b) Explain how the data in Fig. 9.1 and Fig. 9.2 provide evidence for:

(i) allopatric speciation [2]

1. The Carribean islands are geographically isolated* as they are surrounded
by water that acts as a physical barrier preventing interbreeding. This results
in the disruption of gene flow*;
2. The islands due to their differing habitats / environments, present many
different niches, such as different tree types, for species to fill
3. Thus (common) ancestral species on different islands were exposed to
different selection pressures* and natural selection act on them, resulting
in different islands having different mix of species

(ii) sympatric speciation [2]

1. There exist variation such as size or toe lengths in the various lizard
populations and those with favourable traits have a selective advantage to the
local conditions on each island will be selected for, increasing frequency of
favourable alleles / and will survive, reproduce and pass on their alleles to the
next generation;
2. As different sub populations evolved independently of each other, their allele
frequencies changed as they accumulate different genetic mutations, and
were subjected to genetic drift and natural selection.
3. Over a hundreds and thousands of generations, each population on the
different islands became reproductively isolated* and thus evolving into
separate species on each island.

2019 H2 Biology A level Paper 3 [Turn over

 20

(c) On the smallest Caribbean islands, severe storms can wipe out the existing populations
of Anolis lizards. When small numbers of Anolis lizards recolonise these islands, only
slight changes in leg length occur within their populations over time.

Outline how the evolution of Anolis lizards on the larger Caribbean islands differs from
the evolution of Anolis lizards on the smallest islands. [3]
1. On the smallest islands, the small number of pioneers are not likely to carry all the
alleles present in the source population from the larger Carribean islands.
2. The new population of lizards on the smallest islands will usually be small and more
reproductively isolated as compared with the populations on the larger islands.
3. As a result, there will be reduced genetic variation due to genetic drift in the smaller
islands, and hence less changes in leg lengths will occur in these populations over
time

[Total: 10]

2019 H2 Biology A Level Paper 3

 21

10 <Climate change question>

Kelp is a large seaweed (macroalga) that gross.as a ’kelp forest’ in cool shallow sea water. It
provides an important habitat and source of food for many marine species.

Climate change results in higher sea temperatures that allow sea urchins to invade areas of
kelp forest. Sea urchins are herbivores and graze rock surfaces to which kelp is attached.

Fig. 10.1 shows August sea temperatures in Tasmanian kelp forests from 1950 up to 2010.

Fig. 10.1

Fig. 10.2 shows sea urchin density in relation to August sea temperatures around Tasmania.

Fig. 10.2

(a) Using the data in Fig. 10.1 and Fig. 10.2, explain why the invasion of sea urchins into
Tasmanian kelp forests is a recent phenomenon. [2]

At a large number of different sites around Tasmania, marine biologists have recently recorded
the percentage kelp cover and sea urchin density.

The change in percentage kelp cover with increasing sea urchin density is shown in Fig. 10.3.
The size of each circle represents the frequency of sites with the same percentage kelp cover
and sea urchin density.

2019 H2 Biology A level Paper 3 [Turn over

 22

Fig. 10.3

(b) Using the information provided throughout the whole of this question, describe and
explain the likely impact of climate change on Tasmanian kelp forests. [3]

[Total: 5]

2019 H2 Biology A Level Paper 3

 23

11 Tetracycline is a bacteriostatic antibiotic and penicillin is a bactericidal antibiotic. Fig. 11.1

shows how the number of bacteria per cm3 changes over time in culture media, when incubated
in the presence of each of these antibiotics.

Fig. 11.1

(a) With reference to Fig. 11.1, deduce the difference between the actions of bactericidal
and bacteriostatic antibiotics. [2]

1. From 0 to 5 hours, the number of bacteria per cm3 in sample treated with tetracycline
remained constant at 107 indicating that it is bacteriostatic, preventing further
replication of bacteria by binary fission.

2. From 0 to 5 hours, the number of bacteria per cm3 in sample treated with penicillin
decreases from 107 to 102 indicating that it is bacteriocidal, killing the bacterial thus
reducing the number of bacteria per cm3

(b) Describe the effect of penicillin on the number of bacteria per cm3, as shown in Fig. 11.1.
[2]
1. Treatment with penicillin decreases the number of bacteria per cm3 of sample
2. From 0 to 4 hours, the number of bacteria per cm3 in sample treated with penicillin
decreases from 107 to 102
3. From 4 to 5 hours, the number of bacteria per cm3 plateau/does not decrease further

(c) Name the site of action of penicillin in bacterial cells. [1]

Bacteria peptidoglycan cell wall

[Total: 5]

2019 H2 Biology A level Paper 3 [Turn over

2019 H2 A level Paper 3

Section A
Answer all the questions in this section.

1 Curcumin is a yellow pigment found in the spice turmeric, which is used in curry powder.
Curcumin also has medicinal properties.

(a) Eight plant species are known to have roots that contain curcumin. All eight species are
found in Asia and several of these belong to the genus Curcuma in the ginger family
(Zingiberaceae).

Fig. 1.1 shows the evolutionary relationships for five of the eight species with roots that
contain curcumin. Lines that are not labelled represent other species that do not make
curcumin.

Fig 1.1

(i) Name the term used to describe the organisation of species according to their
evolutionary relationships, as shown in Fig. 1.1. [1]

Phylogeny*

2019 H2 Biology A Level Paper 3

[Turn over
 2

(ii) State how the information needed to construct a diagram such as that shown in
Fig. 1.1 can be obtained through molecular techniques. [4]

1. Amplify DNA sequences of a homologous* gene common to all the species of

organisms being compared via polymerase chain reaction;
2. Sequence the DNA fragments obtained from the different organisms;
3. Align the DNA sequences and calculate/compare the differences between
them;
4. Species that are closely related have more similar nucleotide sequences than
do distantly related species and should be shown to diverge from a more recent
common ancestor in the phylogenetic tree.

(iii) The five species named in Fig. 1.1 all share the characteristic of making curcumin.
These five species have never all been classified together in a single genus.

Explain why the characteristic of making curcumin is not sufficient to place all five
species in the same genus. [2]

1. Compared only 1 characteristic which is insufficient evidence to show that they

belong to the same genus;
2. The 5 species could still be significantly different morphologically compared to
those in the same genus.

R: successful interbreeding to give fertile viable offspring.

(iv) The five species named in Fig. 1.1 occur in just two of the 400 or more families of
flowering plants that exist.

Suggest why the ability to make curcumin is limited to these two families of plants.
[2]

1. The two families must have diverged from a common ancestor which had the
ability to make curcumin;

2. The trait for making curcumin first emerged in this common ancestor.

2019 H2 Biology A Level Paper 3

 3

(b) In vitro experiments show that curcumin decreases the proliferation of cancer cells when
cancer cells are cultured in laboratory apparatus.

In one experiment, the mitotic cell cycle of cancer cells in culture was synchronized so
that they all entered G1 phase at the same time. The cells were then transferred to culture
media containing different concentrations of curcumin. Nine hours later, the percentage
of cells in each phase of interphase was recorded. None of the cells had completed the
G2 phase of interphase.

The results of this experiment are shown in Fig. 1.2.

Fig. 1.2

(i) With reference to Fig. 1.2, describe the effect of curcumin on the percentage of
cancer cells in each phase of interphase nine hours after entering G1 phase. [3]

1. The higher the concentration of curcumin the cells were exposed to, the greater
the percentage of cancer cells still in G1 phase* nine hours later;

2. cite data from graph comparing 2 curcumin concentrations and the percentage
of cancer cells at G1 phase. E.g. 76% of cells at G1 phase in 40 μmol dm-3 curcumin
higher than 20% of cells at G1 phase in 0 μmol dm-3 curcumin.

3. The higher the concentration of curcumin the cells were exposed to, the lower
the percentage of cancer cells in S phase* nine hours later;

4. cite data from graph comparing 2 curcumin concentrations and the percentage
of cancer cells at S phase. E.g. 17% of cells at S phase in 40 μmol dm-3 curcumin
lower than 74% of cells at S phase in 0 μmol dm-3 curcumin.

5. No significant effect of curcumin on the percentage of cancer cells at G2 phase*;

6. Around 6%-9.5% cancer cells across all four different curcumin concentrations
in the culture media.

2019 H2 Biology A level Paper 3 [Turn over

 4

(ii) With reference to Fig. 1.2, suggest how curcumin affects the mitotic cell cycle and
how this could help in the treatment of cancer. [4]

1. Curcumin halts the mitotic cell cycle at the G1 checkpoint;

2. Eg. of possible reasons to halt cell cycle:
cause DNA damage, signal absence of growth factors / nutrients;
3. Cancer cells will be unable to progress in the cell cycle / proceed to S phase;
4. Prevent uncontrolled cell division in cancer cells;

2019 H2 Biology A Level Paper 3

 5

The curcumin molecule can bind to many different molecules in cells, including CDK2, which
regulates the first checkpoint in the cell cycle.

Fig. 1.3 shows some of the many molecular targets of curcumin.

↑ shows that the activity of the named molecule is increased by curcumin.
↓ shows that the activity of the named molecule is decreased by curcumin.

Fig. 1.3

(iii) CDK2 affects the multiplication of cancer cells.

Identify one other molecule in Fig. 1.3 that affects the multiplication of cancer cells
and explain how curcumin’s effect on it could help to treat or prevent cancer. [3]

1. p53*;
2. Curcumin could increase the expression / frequency of transcription of the
p53 gene, resulting in production of more p53 proteins;
3. which will result in increased ability to inhibit cell cycle / repair damaged
DNA / promote apoptosis;

OR

4. growth factors (FGF*/TGFβ1*)/ transcription factor NFκB*

5. bind to growth factors to prevent their binding to receptors;
6. prevent stimulation/progression of the cell cycle leading to reduced cell
division;

R: CDK2*

2019 H2 Biology A level Paper 3 [Turn over

 6

Note: Using the information provided in Fig. 1.4, PhK and TNFα are possible
answers to this question too. However, points 4-6 would still be necessary in order
to link PhK and TNFα to the treatment of cancer.

(c) Curcumin is also useful in the treatment of skin tissue damage as a result of burns.
Normally, scar tissue forms when skin tissue is damaged in this way. Scar tissue can
alter a person’s appearance.

Some of the key events that occur in the formation of scar tissue are shown in Fig. 1.4.

Fig. 1.4

(i) With reference to Fig. 1.3 and Fig. 1.4, explain why curcumin is useful in the
treatment of burns to prevent changes in the appearance of skin. [4]

1. Curcumin reduces the activity of PhK* and TNFα* which are released by
damaged/burned skin;
2. Lower levels of PhK activity will decrease activation of NFκB;
3. which upregulates the expression of genes involved in scar tissue
formation;
4. Lower levels of TNFα activity will decrease attraction of white blood cells
and the release of FGF and TGFβ1 by these cells;
5. reducing fibroblast proliferation and differentiation into scar tissue;

2019 H2 Biology A Level Paper 3

 7

(ii) Curcumin is a non-competitive inhibitor of the enzyme phosphorylase kinase, PhK.

Explain how curcumin interacts with the PhK molecule to decrease its activity. [3]

1. Curcumin binds to site other than active site on PhK;

2. Alters the shape/conformation of the specific* Phk active site*
3. so that active site is no longer complementary* in shape and charge to
substrate, hence substrate cannot bind so the rate of reaction is reduced;

(iii) Explain the roles of kinases (such as PhK) and phosphatases in cell signalling. [4]

1. Kinases catalyse the addition of phosphate* groups from ATP* to a

protein;
2. Kinases activate a large number of molecules resulting in signal
amplification;
3. Phosphatases catalyse the removal of phosphate* groups from proteins
by hydrolysis.
4. Phosphatases inactivate relay molecules so that propagation of the signal
will be inhibited/signal will be terminated.

[Total: 30]

2019 H2 Biology A level Paper 3 [Turn over

 8

2 Lymphoid stem cells are a subset of blood stem cells that give rise to the B lymphocytes and T
lymphocytes of the immune system.

(a) (i) State the level of potency of lymphoid stem cells. [1]

Multipotent*

(ii) Outline two defining features of stem cells. [2]

1. A stem cell is an undifferentiated/unspecialized* cell capable of undergoing

proliferation* and self-renewal*;
2. and retains potential to differentiate* to produce specialized cells upon
receiving appropriate molecular signals*;

Many changes occur during the formation of mature activated B lymphocytes from lymphoid
stem cells. Mature activated B lymphocytes secrete IgG antibodies in response to specific
antigens.

Fig. 2.1 shows a simplified outline of some of the stages that occur during this process, up to
the formation of plasma cells.

Fig. 2.1

2019 H2 Biology A Level Paper 3

 9

(b) Name precisely the genetic process occurring at:

(i) stage 1, to give naïve B lymphocytes capable of recognizing different antigens.
[1]

Somatic recombination*

(ii) stage 2, to give an activated B lymphocyte that now produces IgG antibodies
instead of the membrane-bound IgM antibodies. [1]

Class switching*

(iii) Stage 3, to give sub-clones that vary slightly in their ability to bind antigen X. [1]

Somatic hypermutation*

(c) (i) List the ways in which genetic variation is generated during meiosis, identifying all
the stages of meiosis at which each process occurs. [4]

1. Independent assortment involving the random orientation of

homologous chromosomes* at the equator/metaphase plate* during
metaphase I*;
2. And separation of homologous chromosomes at anaphase I* ultimately
results in different combinations of parental chromosomes in the
gametes;

3. Crossing over * between non-sister chromatids of homologous

chromosomes* occurs at prophase I* at points called chiasmata;
4. Where equivalent portions of these chromatids break and rejoin, resulting
in exchange of genetic material/alleles, hence new combination of alleles
on the chromatid/chromosome;

5. Independent orientation of non-identical sister chromatids of each

chromosome during metaphase II* at the metaphase/equatorial plate &
their subsequent separation during anaphase II*.

(ii) Both the process at stage 1 in Fig. 2.1 and the process of meiosis generate genetic
variation.

State how the genetic variation generated by these two processes is different. [2]

1. Somatic recombination only alters the heavy and light chain gene loci
whereas genetic variation in meiosis can alter any gene locus of the
organism;
2. Somatic recombination involves removing and joining of gene segments
whereas crossing over in meiosis involves exchange of equivalent
segments;
3. Somatic recombination occurs on a single chromosome, whereas crossing
over in meiosis involves non-sister chromatids on a pair of homologous
chromosomes;

AVP

[Total: 12]

2019 H2 Biology A level Paper 3 [Turn over

 10

3 <climate change taken out of syllabus for 2020>

One cause of climate change is the emission of carbon dioxide gas into the atmosphere by
human activity.

Table 3.1 shows the percentage of the world’s global carbon dioxide emissions in 2011 that
came from four countries. It also shows how these countries differed in the size of their
populations and in their economic wealth, as a percentage of the world total.

Table 3.1

percentage
percentage share of percentage share of
contribution to global
country global human global economic
carbon dioxide
population wealth
emissions
China 23 19 15
United States of 14 4 17
America
India 6 17 6
Russian Federation 5 2 4

(a) With reference to Table 3.1, explain how a country’s population size and wealth affects
its percentage contribution to global carbon dioxide emissions. [4]

(b) China, India and the United States of America plant young trees to increase the area of
their land covered by forest. This counteracts part of their carbon dioxide emissions.

Explain, with reference to biochemical details, how planting young trees helps to
counteract carbon dioxide emissions. [4]

[Total: 8]

2019 H2 Biology A Level Paper 3

 11

Section B

Answer one question in this section.

Your answers should be illustrated by large, clearly labeled diagrams, where appropriate.
Your answers must be in continuous prose, where appropriate.
Your answers must be set out in sections (a), (b) etc., as indicated in the question.

4 (a) Describe the structure of the nuclear envelope, including the roles of each of the
constituent biomolecules of cell membranes. [15]

1. The nuclear envelope is a double membrane that is perforated with

nuclear pores*;
2. The outer membrane of the nuclear envelope is continuous with the
rough endoplasmic reticulum;

3. Cell membranes contain phospholipids*, cholesterol* and proteins*;

4. The double membrane is made up of 2 phospholipid bilayers*;

5. Each phospholipids are made up of 2 hydrophobic hydrocarbon tails
/ chains* and a hydrophilic phosphate head* are attached to a
glycerol*;
6. Phosphate heads are charged and hydrophilic and will interact with water
and hydrocarbon tails are non-polar and therefore hydrophobic and
arranged away from aqueous medium;
7. The hydrocarbon tails of phospholipids form hydrophobic interactions
with each other, forming a hydrophobic core* in the phospholipid
bilayer *;
8. The hydrophobic core* of phospholipid bilayer serve as a barrier to
movement of polar, charged and large molecules;
9. As interactions between the phospholipids are weak, phospholipids are
able to move laterally in the bilayer, contributing to the fluidity* of the
membrane;

10. Cholesterol is an amphipathic molecule with hydrophobic* four ringed

structure and a hydrophilic* hydroxyl group;
11. The OH group interacts with the phosphate heads of the phospholipids
while the 4-ring structure forms hydrophobic interactions with the
hydrophobic core of the membrane;
12. Cholesterol in involved in the regulation of membrane fluidity*
13. Cholesterol prevents the membrane from being overly fluid at warmer
temperatures as cholesterol’s rigidity restricts phospholipids’ lateral
movement;
14. Cholesterol prevents the membrane from being overly firm at lower
temperatures as cholesterol prevents the close packing of phospholipids
and hence prevents its solidification/ crystallization;

15. Transport proteins embedded in membranes control the movement of

polar, charged and large molecules that are otherwise unable to diffuse
across the membrane;
16. These proteins are transmembrane* proteins with hydrophobic region
that interact with the hydrophobic core* of the membrane allow them to
be embedded in the membrane;
17. They provide a hydrophilic* channel which allows for the movement of
polar and charged molecules across the membrane;

2019 H2 Biology A level Paper 3 [Turn over

 12

18. In the nuclear envelope, nuclear pores are formed by proteins that make
up a nuclear pore complex;
19. Nuclear pores allow for the import of molecules such as ribonucleoside
triphosphates into the nucleus, and the export of molecules such as the
mature mRNA out of the nucleus;

(b) Explain the roles of the nuclear envelope in the functioning of a eukaryotic cell. [10]
1. Nuclear envelope encloses the cell’s genome / DNA and protects the
genome from reactions that are occurring in the cell;
2. It allows for compartmentalisation to occur, spatially separating
transcription* in the nucleus from translation* in the cytoplasm;
3. This allows the formation of unique environments for these processes
involving enzymatic reactions reactions;
4. The nuclear envelope serves as a barrier to large, polar and charged
molecules, allowing only the selected molecules to move in and out of
the nucleus via the nuclear pore*;
5. This allows for localisation of enzymes and substrates in the same
compartment so that reactions can take place more efficiently;
6. The nuclear pores in the nuclear envelope allow for the diffusion of
ribonucleoside triphosphate and the import of enzymes/proteins such as
RNA polymerase* into the nucleus to allow for transcription*;
7. Nuclear pore complexes recognises the 3’ poly-A tail* and the 5’-
methylguanosine cap* on mature mRNA*,
8. ensuring that only mature mRNA is exported out of the nucleus for
translation* in the cytoplasm;
9. The nuclear pores also allow for the import of ribosomal proteins*;
10. so that ribosomal proteins and rRNA* can assemble to form ribosomal
subunits* in the nucleolus*;
11. Assembled ribosomal subunits are then recognised for export* out of the
nucleus into the cytoplasm;

[Total: 25]

5 (a) Many different polymers are formed in cells. A polymer is a chain of monomers.
Describe how different polymers form in cells. [15]

1. Condensation reaction formation of bonds with removal of a water

molecule;
2. Hydrolysis reaction breaking of bonds with the use of a water
molecule;

A. Carbohydrates
3. Starch is made from condensation* reaction of many α-glucose*
monomers;
4. Amylose* - glucose monomers are linked by α(1-4) glycosidic* bonds,
forming a helical structure;
5. Amylopectin* - glucose monomers are linked by α(1-4) glycosidic
bonds within a branch and α(1-6) glycosidic* bonds at branch points,

6. Cellulose is made from condensation* reaction of many β-glucose*

monomers;
7. β-glucose monomers are joined by β(1-4) glycosidic bond* alternate
monomers are inverted / monomers are inverted with respect to one
another,

B. Proteins
8. A specific amino acid is covalently attached to 3’CCA* stem of a tRNA

2019 H2 Biology A Level Paper 3

 13

with a specific anticodon* to form aminocyl-tRNA. This attachment is

catalysed by aminoacyl-tRNA synthetase*.
9. Translation initiation factors will facilitate binding of small ribosomal
subunit and initiator tRNA carrying methionine* to newly synthesized
mRNA strand.
10. The anticodon (UAC) of initiator tRNA will complementary base pair with
start codon (AUG)* of mRNA.
11. Binding of large ribosomal subunit will complete ribosome forming
translational initiation complex.
12. This positions initiator tRNA at peptidyl site (P site)* leaving aminoacyl
site (A site)* vacant for incoming aminoacyl-tRNA molecules.
13. A second aminoacyl-tRNA with a specific anticodon* and corresponding
amino acid, binds to specific mRNA codon at A site via complementary
base-pairing*.
14. A peptide bond* is formed between adjacent amino acids, catalysed by
peptidyl transferase* of large ribosomal subunit. Methionine
dissociates from initiator tRNA as a result and remains bound to second
amino acid at A site.
15. Amino acids undergo condensation* to form polypeptides with the
removal of 1 water molecule;
16. The ribosome translocates in 5’ to 3’* direction, shifting first tRNA to exit
site (E site) allowing it to be released into cytosol. tRNA with growing
polypeptide chain is now at P site and A site will hold a new incoming
aminoacyl-tRNA with an anticodon complementary to next codon on
mRNA.
17. This process continues until a stop codon (UAA/ UAG/ UGA)* is
reached at A site. Release factors* will enter A site causing hydrolysis
of bond between polypeptide chain and tRNA in P site.

C. Nucleic Acids
18. Replication of DNA is semi-conservative* where the original strands of
double helix separates and act as templates* for synthesis of two new
strands;
19. This gives rise to two new DNA molecules, each consisting of one
original and one newly synthesised strand;
20. replication of DNA begins at origin of replication* ;
21. where enzyme helicase* will bind and unzip* DNA molecule by breaking
hydrogen bonds* between complementary base pairs;
22. separated strands of DNA interact with single stranded DNA binding
proteins* so that it will remain single stranded and can serve as a
template* for replication;
23. Topoisomerase* relieves overwinding strain ahead of replication forks
by breaking, swivelling and rejoining DNA strands;
24. enzyme primase* catalyses synthesis of a short RNA primer*; (Reject:
RNA primase)
25. Complementary base pairing* occurs between template* strand and
free incoming deoxyribonucleoside triphosphates*;
26. Whereby adenine* forms 2 hydrogen bonds* with thymine* and
guanine* forms 3 hydrogen bonds with cytosine*;
27. DNA polymerase* catalyses formation of phosphodiester bond*
linking DNA nucleotides;
28. and new DNA strand is synthesized in 5’ to 3’ direction*; (A: template
strand read from 3’ 5’ direction*)
29. RNA primers* will then be removed and replaced by DNA by another
DNA polymerase;

2019 H2 Biology A level Paper 3 [Turn over

 14

30. one of daughter strands known as leading strand* will be synthesised

continuously towards replication fork;
31. other strand known as lagging strand* is synthesised discontinuously
away from replication fork, giving rise to Okazaki fragments*;
32. DNA ligase* catalyses formation of phosphodiester bond* between
Okazaki fragments, sealing the nicks*;
33. RNA polymerase* will attach to promoter* region of a gene on DNA
molecule with aid of protein factors;
34. RNA polymerase will then unzip* DNA double helix by breaking
hydrogen bonds* between complementary base pairs*;
35. 3’ to 5’ DNA strand / one of the DNA strands will be used as the
template* strand for synthesis of a complementary mRNA strand;
36. Free ribonucleotides* will bind by complementary base pairing* to
nucleotides on DNA template strand;
37. where adenine* forms 2 hydrogen bonds* with uracil*, thymine* forms 2
hydrogen bonds* with adenine*, cytosine* forms 3 hydrogen bonds*
with guanine*;
38. RNA polymerase will catalyse formation of phosphodiester bonds*
between free ribonucleotides to form sugar phosphate backbone*;
39. Polymerisation of ribonucleotides will result in formation of a new mRNA
strand that is synthesized in 5’ to 3’ direction*;
40. RNA polymerase will dissociate from template DNA strand when it
reaches termination sequence*;

(b) Explain why enzymes and transmembrane receptors must be made from a [10]
hundred monomers or more in order to fulfill their functions.

1. Enzymes and transmembrane receptors need to have multiple

domains/regions that serve different functions.

Enzymes
2. Active site* contains numerous contact* and catalytic* residues.
3. Contact residues determine enzyme-substrate specificity.
4. Contact/binding residues bind reversibly with the substrate* while
positioning it in the correct orientation.
5. The substrate is held in active site* by weak interactions like hydrogen
bonds* and ionic bonds*.
6. Catalytic residues act on the bonds in the substrate molecule and the
R-groups* of a few of the amino acids residues catalyse the conversion
of the substrate to product.
7. Longer polypeptide chain allows for the formation of allosteric sites e.g.
for inhibitor/activator allow regulation of enzyme activity.
8. Longer polypeptide chain allows for the formation of sites under than
the active site for non-competitive inhibitors to bind to decrease enzyme
activity.
9. Tertiary/quaternary structure* of the enzyme is maintained via
hydrogen bonds*, ionic bonds*, hydrophobic interactions* and
disulfide linkages* which are formed between the R groups* of the
many different amino acids;
10. Structural residues interact to maintain an active site* which is
complementary in shape and charge to the substrate.

2019 H2 Biology A Level Paper 3

 15

11. Large numbers of monomers allow the polypeptide to fold in a globular*

structure, where polar R groups are exposed to water molecules in the
aqueous environment.
12. This allows the enzyme to be soluble since these polar groups can form
hydrogen bonds* with water molecules.

Transmembrane proteins

13. Transmembrane proteins have sufficient non-polar R groups of amino

acids on its exterior surface that can form hydrophobic interactions*
with the non-polar hydrocarbon chains of the phospholipids in the
bilayer;

14. The charged phosphate heads of the phospholipid bilayer interact with
the charged / polar R groups of amino acids found on the exterior
surface of the channel protein;

15. This allows the proteins to span the membrane;

16. In channel proteins/carrier proteins, the amino acid residues can also
form a hydrophilic* channel/binding site for the passage of polar* or
charged* molecules through the hydrophobic* core of the membrane.

17. In receptor proteins, the residues form a ligand-binding site which allows
an extracellular signal to give rise to an intracellular response.

Total: 25]

---- End of Paper -----

2019 H2 Biology A level Paper 3 [Turn over