CHAPTER > 11

Biotechnology :
Principles and
Processes
NEET KEY NOTES
Å Biotechnology is the technique of using living organisms or Å Techniques of genetic engineering include construction of
enzymes from organisms to produce products and processes recombinant DNA, gene cloning and gene transfer. These
useful to humans. techniques allow the isolation and introduction of a set of
Å Biotechnology deals with the large scale production and desirable genes without introducing undesirable genes into
marketing of products such as enzymes, insulin or the target organism.
antibiotics, etc., that are of importance to mankind. Å Origin of replication (ori) is a specific DNA sequence in the
Å The European Federation of Biotechnology (EFB) has given chromosome which can initiate DNA replication. The foreign
a definition of biotechnology as, the integration of natural DNA introduced into the host genome has to be linked the
science and organisms cells, parts thereof and molecular origin of replication in the host chromosome for the gene to
analogues for products and services’. be able to multiply. This is also known as cloning which
involves making multiple identical copies of any template
Principles of Biotechnology DNA. If the foreign gene is not linked to the ori sequence it
may not be able to multiply.
Following two core techniques gave birth to modern
Å In 1972, the first recombinant DNA was constructed by
biotechnology
Stanley Cohen and Herbert Boyer. They isolated the
Å Genetic engineering It is the alteration of the chemistry of
antibiotic resistance gene by cutting out a piece of DNA from
genetic material (DNA/RNA), introduce these into host a plasmid (autonomously replicating circular extra-
organisms and consequently change the phenotype of the chromosomal DNA) of Salmonella typhimurium. The cutting
host organism. of DNA at specific locations become possible with the help
Å Bioprocess engineering It is the maintenance of sterile
of restriction enzymes (molecular scissors).
conditions in order to enable the growth of only desired Å The cut pieces of DNA were then linked with the plasmid
microbes or eukaryotic cells in large quantities for the DNA using DNA ligase enzyme. These plasmids act as
production of antibiotics, enzymes, hormones, vaccines, etc. vectors to transfer the piece of DNA attached to it into the
host organism. This makes a new circular autonomously
Genetic Engineering replicating DNA created in vitro and is known as the
Å It is the deliberate modification of an organism’s DNA, using recombinant DNA.
various techniques. This altered DNA (recombinant DNA) is Å The recombinant DNA is transferred into Escherichia coli, a
then introduced into host organisms to change their bacterium closely related to Salmonella, where it replicates
phenotype. using the new host’s DNA polymerase enzyme and makes
Å This is followed by growing this genetically modified cell in multiple copies of itself. It also produces multiple copies of
large quantities, by maintaining sterile environment, for the the antibiotic resistance gene in the new host (E. coli). This
manufacture of biotechnological products like antibiotics, process is called as cloning of antibiotic resistance gene in
vaccines, enzymes, etc. E. coli.
Å There are three basic steps involved in genetically Types of Restriction Enzyme
modifying an organism Å The restriction enzymes can be of three types, on the basis
n
Identification of DNA with desirable genes. of their chemical and physiological properties.
n
Introduction of the identified DNA into the host.
Type II Type III
Maintenance of introduced DNA in the host and Features Type I Enzyme
n
Enzyme Enzyme
transfer of the DNA to its progeny. Protein Bifunctional enzyme Separate Bifunctional
structure with 3 subunits endonuclease enzyme with
Tools of Recombinant DNA and methylase 2 subunits
Recognition Bipartite and Short sequence Asymmetrical
Technology site asymmetrical (4-6 bp), often sequence of 5-7
(e.g. TGAC palindromic bp
Genetic engineering or recombinant DNA technology can be and TGCT)
accomplished only if we have the key tools, i.e. restriction
Cleavage site Non-specific >1000 bp Same as or 24-26 bp
enzymes, polymerase enzymes, ligases, vectors and the host from recognition site close to downstream of
organism. recognition site recognition site
Restriction and Mutually exclusive Separate Simultaneous
1. Restriction Enzymes methylation reactions
Å In the year 1963, the two enzymes responsible for restricting ATP needed Yes No Yes
the growth to bacteriophage in E. coli were isolated. One of for restriction
these added methyl group to DNA, while the other cut Mg 2+needed Yes Yes Yes
DNA. The latter was called restriction endonuclease. for restriction
Å The first restriction endonuclease, i.e. Hind II was isolated Commonly Random cutting and Gene cloning Gene cloning
used in fragments making
by Smith Wilcox and Kelley in 1968.
Hind II always cut the DNA at specific base sequences, i.e. Examples of Restriction Enzymes
of six base pairs. Apart from Hind II, more than Names Sources Sites Types of End
900 restriction enzymes have been isolated from over
230 strains of bacteria. Hpa I Haemophilus 5 ′ GTT - AAC 3 ′ Blunt
parainfluenzae 3 ′ CAA - TTG 5 ′
Å Naming of restriction enzyme proceeds in a way that the
Ssp I Sphaerotilus 5 ′ AAT - ATT 3 ′ Blunt
first letter of the name comes from the genus and the species 3 ′ TTA - TAA 5 ′
second two letters come from the species of prokaryotic
cell. Roman number following the names indicate the order Pst I Providencia 5 ′ CTGCA - G 3 ′ Sticky
in which the enzyme were isolated from that strain of stuartii 3 ′ G - ACGTC 5 ′
bacteria, e.g. Eco RI comes from E. coli RY13. Hind II Haemophilus 5 ′ GTC - GAC 3 ′ Blunt
influenzae 3 ′ CAG - CTG 5 ′
Å Restriction enzymes belong to a larger class of enzymes
called nucleases, which are of the following two types Eco RI Escherichia coli 5 ′ G - AATTC 3 ′ Sticky
3 ′ CTTAA - G 5 ′
n
Exonucleases, which remove nucleotides from the ends
of the DNA (either 5′ or 3′) in one strand of duplex. Hae III Haemophilus 5 ′ GG - CC 3 ′ Blunt
aegyptius 3 ′ CC - GG 5 ′
n
Endonucleases make cuts at specific positions within
the DNA by inspecting the length of a DNA sequence. Bam HI Bacillus 5 ′ GGAT - CC3 ′ Sticky
Once finds its specific recognition sequence, it will bind amyloliquefaciens 3 ′ CCTA - GG 5 ′
to the DNA and cut each of the two strands of the
double helix at specific points in their sugar phosphate Å Restriction enzymes cut the strand of DNA, a little away
to backbones. from the center of the palindrome sites, but between the
Å These restriction enzymes recognise a palindromic same two bases on the opposite strands. These staggered
nucleotide sequence in the DNA and cut both the strands cuts leave single-stranded portions at both the ends.
of DNA at that point. Palindrome in the DNA is a sequence These are referred to as sticky ends.
of base pairs that reads same on the two strands when Å There are other restriction enzymes which cut both the
orientation of reading is kept the same. DNA strands at the same place so that single-stranded
Å For example, the following sequence reads the same on the pieces are not left in the ends. Such ends are called blunt
two strands whether read in 5′→ 3′ direction or 3′→ 5′ ends.
direction. Å Stickiness is the chemical ability of a DNA molecule to
5′−GAATTC−3′ base pair with any other DNA molecule that has also been
3′−CTTAAG−5′ cut by the same restriction enzyme. It means it will have

NEET KEY NOTES

 same sequence hanging unpaired. This stickiness of the Å The DNA fragments separated and cut out from the gel and
ends facilitates the action of the enzyme DNA ligase. extracted from gel piece. This step is known as elution.
DNA ligase (molecular binder) enzymes help in sealing
Å

the gaps in DNA fragments by forming a phosphodiester

2. Cloning Vectors
bond between the adjacent 3′ — OH and 5′ phosphate Å Plasmids and bacteriophages are used as cloning vectors.
terminals, thereby joining nicks in double-stranded DNA. This is because plasmids and bacteriophages have the
ability to replicate within the bacterial cell independent of
Å Lysing enzymes/Lyases enzymes are used for the isolation chromosomal DNA.
of DNA from cells, e.g. lysozyme is used to digest the Å Bacteriophages have very high copy numbers of their
bacterial cell wall for the extraction of cellular DNA.
genome within the bacterial cells. But in case of plasmids,
Protease, lipase and other degrading enzymes come in this
some may have only one or two copies per cell whereas
category. others may have 15-100 copies per cell.
Å Alkaline phosphatase catalyse the removal of 5′ Å Cloning vectors use the machinery of bacterial cell to
phosphate group from the DNA and thus, modify the replicate and thereby, increase the copy number (make
terminus of DNA. clones) of the DNA inserted into them.
Action of restriction enzyme Å These help easy linking of foreign DNA and allow the
The enzyme cuts both DNA Eco RI cuts the DNA between bases selection of recombinants (bacterial cells that have picked
strands at the same site G and A only when the sequence
GAATTC is present in the DNA
up recombinant plasmid) from non-recombinants (those
Vector DNA Foreign DNA who have not).
Å All vectros have four special features that are required to
facilitate cloning into a vector.
Å Origin of replication (ori) is the sequence from where
Eco RI replication starts and any piece of DNA when linked to
Sticky end
this sequence can be made to replicate within the host
cells. This sequence also controls the copy number of the
linked DNA.
Sticky end
Å Selectable marker helps in identifying and eliminating
DNA fragments join at sticky ends
non-transformants and selectively permitting the growth
of the transformants.
n
Transformation is the procedure through which a piece
of DNA is introduced into a host bacterium.
Recombinant DNA
Steps in the formation of recombinant DNA by action of restriction
n
Normally, antibiotics such as ampicillin,
endonuclease enzyme Eco RI chloramphenicol, tetracycline or kanamycin, etc., are
considered useful selectable markers.
Separation and Isolation of DNA Å Cloning sites are used to link the alien DNA. The vector
Fragments needs to have cloning or recognition sites for the
commonly used restriction enzymes. The presence of more
Å Separation of restriction fragments from each other can be than one recognition sites within the vector will generate
done by gel electrophoresis. several fragments and complicate the gene cloning.
Å An electric field is applied and these fragments are forced Cla I Hind III
Eco RI
to move through a viscous gel of agarose (a natural
polymer extracted from sea weeds). Pvu I
Bam HI
Pst I
Å Since, DNA fragments are negatively charged (because of ampR
their phosphate groups) they will move towards the tetR
pBR322 Sal I
positively charged pole, i.e. anode.
Å Smaller fragments move faster and bigger ones move ori rop
slowes, but they all separate out (according to their size)
into bands that can be identified later by staining.
Pvu II
Å The staining of DNA bands can be done with a compound
E. coli cloning vector pBR322 showing restriction sites (Hind III,
known as ethidium bromide followed by exposure to UV EcoRI, BamHI, Sal I, Pvu II, Pst I, Cla I), ori and antibiotic resistance
radiation. Bright orange coloured bands of DNA are genes (amp R and tet R ). rop codes for the proteins involved in the
visible. replication of the plasmid

NEET KEY NOTES

 n
The ligation of foreign DNA is carried out at a restriction Å Recombinant DNA is directly injected into the nucleus of
site present in one of the two antibiotic resistance an animal cell, using a microsyringe. This is known as
genes. It can be done at the Bam HI site of tetracycline microinjection.
resistance gene in the vector pBR 322. Å Plant cells are bombarded with high velocity
n
In this case, the recombinant plasmids will lose microparticles of gold or tungsten coated with DNA in a
tetracycline resistance due to the insertion of foreign method known as biolistics or gene gun.
DNA but can still be selected out from non-recombinant Å Some pathogens that naturally infect a cell can be
ones by plating the transformants on a medium ‘disarmed’ (by eliminating their harmful gene) and then
containing ampicillin. allowed to infect the cell, carrying the desired
n
The recombinants will grow in ampicillin containing recombinant DNA into the host.
medium but not on that containing tetracycline. But,
non-recombinants will grow on the medium containing
both the antibiotics.
Processes of Recombinant
n
Other selectable markers can differentiate recombinants DNA Technology
from non-recombinants on the basis of their ability to Å Recombinant DNA technology involves various steps in a
produce colour in the presence of a chromogenic specific sequence such as isolation of the desired genetic
substrate. In this, recombinant DNA is inserted within material (DNA), cutting of DNA at specific locations,
the coding sequence of an enzyme, β-galactosidase.
isolation of desired DNA fragment, amplification of gene
This results into inactivation of the gene synthesis of this
of interest by PCR, ligation of DNA fragments into a
enzyme and referred to as insertional inactivation.
vector, insertion of recombinant DNA into host cell,
n
The presence of a chromogenic substrate gives blue
culturing the host cells in a medium at large scale and
coloured colonies if the plasmid in the bacteria does not
interaction of the desired product.
have an insert and if the colonies have inserted plasmid,
such colonies are recombinants and show no Isolation of the Genetic Material
colouration.
Å Vectors for cloning genes in plants and animals A
(DNA)
pathogen of several dicot plant is able to deliver a piece of Å The genetic material, DNA needs to be in pure form, free
DNA, i.e. ‘T-DNA’ to transform normal plant cells into a from other macromolecules, for restriction enzymes to be
tumour cells and disect these tumour cells to produce the able to act on it. Following are the steps
chemicals required by the pathogen. Similarly, retrovirus in n
Cell wall needs to be broken to release DNA. For this,
animals have the ability to transform normal cells into the bacterial cells/plant or animal tissue are treated
cancerous cells. with enzymes such as lysozyme (bacteria), cellulase
Å The tumour inducing (Ti) plasmid of Agrobacterium (plant cells), chitinase (fungus).
tumefaciens has now been modified into a cloning vector n
Histones and other proteins have to be removed by
which is able to use the mechanisms to deliver genes of our treatment with proteases and the RNA can be removed
interest into a variety of plants. by treatment with ribonuclease.
Å Similarly, retroviruses can be disarmed and used to deliver n
With the addition of chilled ethanol, purified DNA
desirable genes into animal cells. precipitates out. This can be seen as collection of fine
threads in the suspension and removed out by
3. Competent Host spooling.
Å Since, DNA is a hydrophilic molecule, it cannot pass
through cell membrane that has hydrophobic ends both on Amplification of Gene of Interest
the inside and outside. Hence, various artificial means Using PCR
have to be used to make the cells competent to take up Å PCR stands for Polymerase Chain Reaction, a method of
foreign DNA.
amplifying the fragments of DNA. This method can make
Å Bacteria are treated with a specific concentration of a multiple copies of even a single DNA fragment or the
divalent cation, such as calcium, which makes the cell gene of interest in a test tube. The reaction mixture
membrane more permeable. Recombinant DNA can then be requires
forced into such cells by incubating the cells with n
Double-stranded DNA fragment (gene of interest).
recombinant DNA on ice, followed by placing them briefly
at 42°C (heat shock) and then putting them back on ice.
n
Primers, i.e. small chemially synthesised
This makes the bacterial cell competent to take up the oligonucleotides that are complementary to the
recombinant DNA. regions of this DNA.

NEET KEY NOTES

 n
The special thermostable DNA polymerase (isolated Obtaining the Foreign Gene
from the bacterium, Thermus aquaticus) that does not
denature and remains active even at high temperatures. Product
Å With each round of reaction, the DNA quantity gets Å The foreign gene gets expressed under appropriate
amplified. If the process of replication of DNA is repeated conditions. After having the cloned gene of interest and
many times the segment of DNA can be amplified to having optimised conditions to induce the expression of
approximately billion times, i.e. 1 billion copies are made. the target protein, large scale production can be carried
out.
Region to be amplified Å If any protein encoding gene is expressed in a
heterologous host, the product protein is called a
5′ 3′ dsDNA
3′ 5′ recombinant protein.
Heat Denaturation Å The cells harbouring cloned genes of interest may be
5′ 3′ grown on a small scale in the laboratory. The cultures
3′ 5′ may be used for extracting the desired protein. The cells
Primers annealing
5′ 3′ can also be multiplied in a continuous culture system,
3′ 5′ wherein the used medium is drained out from one side,
DNA polymerase
(Taq polymerase while fresh medium is added from the other to maintain
+ deoxynucleotides) the cells in their physiologically most active growth
5′ 3′ phase, i.e. log/exponential phase. This type of culturing
3′ 5′ Extension method produces a larger biomass leading to higher
5′ 3′
3′ 5′ yields of desired protein. For large scale production,
bioreactors are used and culture can be processed.
30 cycles
Å Bioreactors are large vessels (100-1000 L capacity) in
Amplified which raw materials are biologically converted into
(~ 1 billion times) specific products, individual enzymes, etc., using
microbial, plant, animal or human cells.
Polymerase Chain Reaction (PCR) : Each cycle has three
steps, i.e. These are specially designed to provide optimal
conditions of temperature, pH, substrate, salts, vitamins,
oxygen, etc., for achieving the desired production levels.
Insertion of Recombinant DNA into Å Bioreactors can be of two types–continuous culture
the Host Cell/Organism system and non-continuous culture system (Batch
culture). The two variants of bioreactors are sparged
Å There are several methods of introducing the ligated DNA tank bioreactor and simple stirred-tank bioreactor. A
into recipient cells. Recipient cells after making them stirred tank reactor is usually cylindrical or with a
competent to receive take up DNA present in their curved base to facilitate the even mixing and oxygen
surrounding. availability throughout the bioreactor. It is the most
Å So, if a recombinant DNA bearing gene for resistance to an commonly used type of bioreactor.
antibiotic (e.g. ampicillin) is transferred into E. coli cells, the
host cells are transformed into ampicillin-resistant cells. Downstream Processing
Å If we spread the transformed cells on agar plates containing The steps of downstream processing are separation and
ampicillin, only transformants will grow and untransformed purification of the gene product, i.e. the functional protein,
recipient cells will die. formulating it with suitable formulation. Such
Å Since, due to ampicillin resistance gene, one is able to select a formulations undergo clinical trials to determine product
transformed cell in the presence of ampicillin. The ampicillin quality. The downstream processing and quality control
resistance gene in this case is called a selectable marker. testing vary from product to product.

NEET KEY NOTES

266 Master The NCERT > BIOLOGY (Vol-II)

Mastering NCERT
MULTIPLE CHOICE QUESTIONS

TOPIC 1 ~ Principles of Biotechnology and Restriction Enzymes

1 The controlled use of biological agents, such as live 9 The construction of the first recombinant DNA was
organisms or enzymes from organisms to produce done by using the native plasmid of
products and processes useful to humans is called as (a) E. coli (b) Salmonella typhimurium
(a) biochemistry (b) molecular biology (c) Bacillus thuringiensis (d) Yeast
(c) biotechnology (d) microbiology 10 The first recombinant DNA was constructed by
2 The techniques/processes that are included under (a) Stanley Cohen (b) Herbert Boyer
biotechnology are (c) Temin and Baltimore (d) Both (a) and (b)
(a) in vitro fertilisation (b) correcting a defective gene 11 The linking of antibiotic resistance gene with the
(c) synthesising a gene (d) All of these plasmid vector became possible with
3 EFB stands for (a) DNA ligase (b) RNA ligase
(a) European Federation of Biotechnology (c) DNA polymerase (d) RNA polymerase
(b) Eurasian Federation of Biotechnology 12 The different basic steps of genetically modifying an
(c) East Asia Federation of Biotechnology organism are given below randomly.
(d) Ethiopian Federation of Biotechnology
I. Identification of DNA with desirable genes.
4 The definition of biotechnology given by EFB II. Transfer of the DNA to its progeny.
encompasses III. Maintenance of introduced DNA in the host.
(a) traditional biotechnology
IV. Introduction of identified DNA into the host.
(b) modern molecular biotechnology
(c) DNA fingerprinting Which of the following represents the correct
(d) Both (a) and (b) sequence of steps?
(a) I, II, III and IV (b) I, IV, III and II
5 The two main techniques that gave birth to modern
(c) III, IV, II and I (d) I, III, IV and II
biotechnology are
I. bioprocess engineering. 13 The key tools required for the recombinant DNA
II. genetic engineering. technology are
III. human genome engineering. I. restriction enzymes II. polymerase enzymes
IV. molecular biology. III. ligases IV. vector
Choose the correct option. V. host organism
(a) I and II (b) I and III (c) II and IV (d) II and III Select the correct option.
6 Genetic engineering techniques include (a) I, II and III (b) I, III, IV and V
(a) altering genetic material (c) I, II, III and V (d) I, II, III, IV and V
(b) sequencing genetic material 14 The enzymes, commonly used in genetic engineering
(c) studying genetic material are
(d) None of the above (a) restriction endonuclease and polymerase
7 The specific sequence of DNA that initiate replication (b) endonuclease and ligase
of alien DNA in rDNA technology is called as (c) restriction endonuclease and ligase
(a) initiation sequence (b) origin of replication (d) ligase and polymerase
(c) origin of DNA (d) initiation of DNA 15 The first restriction endonuclease to be discovered is
8 Autonomously replicating circular (a) Hind III (b) Hind II (c) Eco RI (d) Eco RII
extra-chromosomal DNA is 16 Which of the following is a restriction endonuclease?
(a) vector (b) capsid CBSE-AIPMT 2015
(c) plasmid (d) bacteriophage (a) Protease (b) DNase I (c) RNase (d) Hind II
17 The restriction enzyme responsible for the cleavage of 27 Restriction endonuclease enzymes are used to cut
following sequence is (a) single-stranded RNA (b) double-stranded DNA
5′ – G T C G A C 3′ (c) single-stranded DNA (d) double-stranded RNA
3′ – C A G C T G 5′ 28 Restriction endonuclease binds to DNA and cuts two
(a) Alu I (b) Bam HI (c) Hind II (d) Eco RI strands of double helix at specific points in their
(a) sugar-phosphate backbone
18 How many restriction enzymes are isolated till now? (b) hydrogen bond
(a) 920 (b) 940 (c) 900 (d) 230 (c) glycosidic bonds
19 Number of bacterial strains from which restriction (d) None of the above
enzymes have been isolated. 29 Special sequence in the DNA recognised by
(a) 230 (b) 250 (c) 200 (d) 220 restriction endonuclease is called
20 In the naming of restriction enzymes, the first letter of (a) restriction nucleotide sequence
the name is derived from ..... A .... and next two letters (b) palindromic nucleotide sequence
from the ..... B ..... and fourth letters from the name of (c) recognition nucleotide sequence
..... C .... of .... D .... from which the enzymes are (d) All of the above
extracted. 30 Palindrome sequences
A to D in the statement can be (a) read opposite on two strands
A B C D (b) read specific sequence in opposite direction
(a) genus species strain bacteria (c) read same on two strands when orientation of reading is
(b) species genus strain bacteria same
(c) genus species variety eukaryote (d) read opposite on two strands when orientation of
(d) species genus variety eukaryote reading is same
21 There is a restriction endonuclease called Eco RI. 31 Restriction enzyme cuts the DNA strand a little away
What does ‘co’ part in it stands for? from the centre of palindrome site between
(a) Coelom (b) Strain of bacterium (a) same two bases on same strand
(c) coli (d) Colon (b) same two bases on opposite strand
22 The Roman number following the name of restriction (c) opposite bases on same strand
enzyme indicate (d) opposite bases on opposite strand
(a) order in which enzyme is isolated from strain of bacteria 32 The foreign DNA and the vector is cut with the
(b) number of enzyme (a) two different enzymes
(c) order of enzyme (b) same restriction enzymes
(d) None of the above (c) DNA ligase
23 Restriction enzyme belongs to which class of (d) Both (a) and (b)
enzymes? 33 How many fragments will be generated, if a linear
(a) Ligases (b) Exonucleases DNA molecule is digested with a restriction enzyme
(c) Nucleases (d) Proteases having 4 recognition sites on the DNA?
24 In a genetic engineering experiment, restriction (a) 3 (b) 5
enzymes can be used for (c) 4 (d) 6
(a) bacterial DNA only (b) viral DNA only 34 How many fragments will be generated, if a closed
(c) any DNA fragment (d) eukaryotic DNA only circular DNA molecule is digested using a restriction
25 An enzyme catalysing the removal of nucleotides enzyme having six recognition sites on the DNA?
from ends of DNA is NEET (Odisha) 2019 (a) 4 (b) 6 (c) 7 (d) 5
(a) DNA ligase (b) endonuclease 35 A foreign DNA and plasmid cut by the same
(c) exonuclease (d) protease restriction endouclease can be joined to form a
26 Restriction endonucleases are enzymes which recombinant plasmid using NEET 2016
(a) make cuts at any position within the DNA molecule (a) Eco RI (b) Taq polymerase
(b) recognise a specific nucleotide sequence for binding (c) polymerase III (d) ligase
and then cleaves both the strands of DNA
36 Which is also called molecular glue? JIPMER 2019
(c) restrict the action of the enzyme DNA polymerase
(a) DNA gyrase (b) DNA helicase
(d) remove nucleotides from the ends of the DNA molecule
(c) DNA ligase (d) DNA polymerase
37 Study the given diagram and identify the enzymes A and B (a) A–Restriction endonuclease, B–Restriction
involved in steps I and II. exonuclease, C–RNA ligase, D–Transformation
(b) A–Restriction endonuclease, B–Restriction
endonuclease, C–DNA ligase, D–Transformation
Vector (c) A–Restriction exonuclease, B–Restriction
DNA endonuclease, C–DNA polymerase,
Foreign DNA D–Transduction
Enzyme A
Step I (d) A–Restriction endonuclease, B–Restriction
endonuclease, C–DNA polymerase,
Sticky end D–Transformation
40 The cutting of DNA by ……… results in the
fragments of DNA. Choose the appropriate
Sticky end
Enzyme B option.
Step II DNA fragments join at sticky ends (a) restriction endonucleases
(b) exonuclease
(c) endonuclease
(d) anhydro L-galactose
41 DNA fragments generated by the restriction
Recombinant DNA
endonucleases in a chemical reaction can be
Step I Step II separated by NEET 2013
(a) Eco RI DNA ligase (a) centrifugation
(b) Alu I DNA ligase (b) polymerase chain reaction
(c) Hind II DNA polymerase (c) electrophoresis
(d) Restriction endonuclease DNA polymerase (d) restriction mapping
38 Which of the following option (s) is not correct regarding 42 Which of the following techniques is most
Eco RI enzyme? commonly used to separate DNA molecules by
(a) Restriction endonuclease enzyme size?
(b) Isolated from Escherichia coli RY13 (a) Chromatography (b) PCR
(c) Cuts at specific position within the DNA (c) RFLP (d) Gel electrophoresis
(d) None of the above
43 Agarose is extracted from
39 The flowchart given below represents the process of (a) sea weeds (b) blue-green algae
recombinant technology. Identify A to D in the given (c) Ephedra (d) Sargassam
process. 44 What is the criterion for DNA fragments
movements on agarose gel during gel
Foreign DNA Vector
DNA electrophoresis ? NEET 2017
(plasmid) (a) The larger the fragment size, the farther it moves
(b) The smaller the fragments size, the farther it moves
A (enzyme) B (enzyme) (c) Positively charged fragments move to farther end
(d) Negatively charged fragments do not move
45 In gel electrophoresis, restriction enzyme
digested DNA is loaded in wells near
C (joining enzyme) (a) anode (b) cathode
(c) centre of gel (d) any where in the gel
46 Having become an expert on gel electrophoresis,
Recombinant DNA you are asked to examine a gel for a colleague.
molecule
Where would you find the smallest fragments of
DNA?
D (process)
(a) Near the positive electrode, farthest away from the
wells
E. coli (b) Near the negative electrode, close to the wells
Cells divide
(c) Near the top, near the negative pole
(Cloning host)
(d) Near the middle they tend to slow-down after the
first few minutes
47 The DNA fragments separated on an agarose gel can 49 When the DNA fragments are observed under UV
be visualised after staining with NEET 2017 light, they are seen as
(a) bromophenol blue (a) yellow coloured bands
(b) acetocarmins (b) orange coloured bands
(c) blue coloured bands
(c) aniline blue
(d) Both (a) and (b)
(d) ethidium bromide
50 In gel electrophoresis, the separated bands of DNA
48 In gel electrophoresis, the separated DNA fragments
are cut out and extracted from the gel piece. This step
are visualised after staining the DNA with EtBr
is called
followed by exposure to …. .
(a) elution
Choose the appropriate option. (b) origin of replication
(a) Infrared radiation (b) UV-radiation (c) competency
(c) γ-rays (d) Radiowave (d) transformation

TOPIC 2 ~ Cloning Vectors and Competent Host

51 In recombinant DNA technique, the term vector refers 58 Identify A, B, C and D in the given diagram of E. coli
to a cloning vector pBR322.
(a) donor DNA, it is identified and picked up through
A Cla I Hind III
electrophoresis
(b) plasmid transfers DNA into host cell Pvu I
(c) collection of entire genome in the form of plasmid Pst I B
(d) enzyme, cuts the DNA at specific sites D tetR
52 Which of the following is used in recombinant DNA pBR322 Sal I
technique? C
rop
(a) Cell wall of virus
(b) Gene which produces capsid of virus
(c) Bacteriophage Pvu II
(d) Capsid of virus (a) A–Eco RI, B–Bam HI, C–ori, D–ampR
53 During ‘gene cloning’ which is called a gene taxi? (b) A–ampR, B–ori, C–Bam HI, D–Eco RI
(a) Vaccine (b) Plasmid (c) A–ori, B–Bam HI, C–Eco RI, D–ampR
(c) Bacteria (d) Protozoa
(d) A–Bam HI, B–Eco RI, C–ampR, D–ori
54 Which of the following is not a feature of the
59 The given figure is the diagrammatic representation
plasmids? CBSE-AIPMT 2015
of the vector pBR322. Which one of the given
(a) Circular structure
options correctly identifies its certain component(s)?
(b) Transferable
Eco RI Cla I
(c) Single-stranded Hind III
(d) Independent replication
Pvu I
55 Which vector can clone only a small fragment of Pst I Bam HI
DNA? CBSE-AIPMT 2014
ampR tetR
(a) Bacterial artificial chromosome
pBR322 Sal I
(b) Yeast artificial chromosome
(c) Plasmid ori
(d) Cosmid rop
56 The DNA used as a carrier for transferring a fragment
of foreign DNA into a suitable host is called
Pvu II
(a) cloning vector (b) vehicle DNA
(c) gene carrier (d) All of these (a) ori–original restriction enzymes
57 Which of the following is a plasmid vector? (b) rop–reduced osmotic pressure
(a) pBR322 (b) Bam II (c) Hind III, Eco RI–selectable markers
(c) SaI I (d) Eco RI (d) ampR, tetR–antibiotic resistance genes
60 The two antibiotic resistance genes on vector pBR322 70 Recombinant colonies in insertional inactivation are
are for NEET (Odisha) 2019 differentiated on the basis of
(a) ampicillin and tetracycline (a) production of blue colour
(b) ampicillin and chloramphenicol (b) production of no colour
(c) chloramphenicol and tetracycline (c) production of red colour
(d) tetracycline and kanamycin (d) production of green colour
61 The function of ori in a vector is 71 Agrobacterium tumefaciens delivers a piece of DNA
(a) help in replication of linked DNA into dicot plant. The piece of DNA is called as
(b) control copy number of the linked DNA (a) rDNA (b) T-DNA (c) mDNA (d) cDNA
(c) help in selecting recombinants 72 Retroviruses in animals including humans are able to
(d) Both (a) and (b) change normal cells into
62 A gene, whose expression helps to identify (a) germ cell (b) cancerous cells
transformed cells is known as NEET 2017 (c) cosmid (d) vector
(a) selectable marker (b) vector 73 The plasmid of Agrobacterium tumefaciens that is
(c) plasmid (d) structural gene now modified as a cloning vector is AIIMS 2019
63 A selectable marker is used to NEET (Odisha) 2019 (a) Pi-plasmid (b) cosmid
(a) help in eliminating the non-transformants, so that the (c) Ti-plasmid (d) None of these
transformants can be regenerated
74 In Ti-plasmid, which of the following is removed?
(b) identify the gene for a desired trait in an alien organism
(a) Auxin gene (b) Virulent gene AIIMS 2019
(c) select a suitable vector for tansformation in a specific
(c) Cytokinin gene (d) Auxin and cytokinin gene
crop
(d) mark a gene on a chromosome for isolation using 75 Why foreign DNA cannot pass through cell
restriction enzyme membrane?
64 If recombinant DNA carrying antibiotic resistance (a) DNA is hydrophobic
gene (e.g. ampicillin) is transferred into E. coli cell, the (b) DNA is hydrophilic
host cell is transformed into ampicillin resistant cells. (c) DNA is rich in proteins
The ampicillin resistant gene in this case is called a (d) DNA is heavy
(a) vectors (b) plasmid 76 The treatment of host cell with divalent cation leads to
(c) selectable marker (d) cloning sites the
65 The presence of more than one recognition site within (a) change in permeability of DNA
vector will lead to the (b) increased efficiency with which DNA enters the
(a) generation of several fragments bacterium
(b) generation of one fragment (c) decreased efficiency with which DNA enters the
bacterium
(c) generation of half fragment
(d) change in permeability of host
(d) None of the above
66 The recognition site for Bam HI in pBR322 is present in 77 The method which is used to introduce recombinant
(a) ampicillin resistant site (b) tetracycline resistant site DNA into animal cell?
(c) ori site (d) rop site (a) Gene gun method (b) Changing permeability of host
(c) Biolistic method (d) Microinjection
67 When an alien DNA is ligated in tetracycline resistant
gene, the recombinant 78 Which of the following methods(s) is used to
(a) become tetracycline resistant introduce foreign DNA into plant host cells?
(b) will loose tetracycline resistant (a) Gene gun method (b) Gel electrophoresis
(c) will remain same (c) Elution (d) Extension
(d) None of the above 79 For transformation, microparticles coated with DNA
68 The method(s) that is/are used to differentiate to be bombarded with gene gun are made up of
recombinants and non-recombinants is/are CBSE-AIPMT 2012
(a) antibiotic affected gene (b) insertional inactivation (a) silver or platinum (b) platinum or zinc
(c) gene cloning (d) Both (a) and (b) (c) silicon or platinum (d) gold or tungsten
69 In insertional inactivation, the recombinant DNA is 80 DNA transfer with high velocity micro particles is
inserted within the coding sequence of present in JIPMER 2018
(a) β-galactosidase (b) tetracycline resistant gene (a) biolistics (b) hybridisation
(c) restriction enzyme (d) ampicillin resistant gene (c) tissue culture (d) vegetative propagation
CHAPTER 11 > Biotechnology : Principles and Processes 271

TOPIC 3 ~ Processes of Recombinant DNA Technology

81 The different steps involved in the process of 89 Primers are
recombinant DNA technology are given below (a) small chemically synthesised oligonucleotides of about
randomly? Arrange these in correct order. 10-18 nucleotides that are complementary to the region
I. Extraction of the desired gene product. of template DNA
(b) chemically synthesised oligonucleotides of about 10-18
II. Amplification of the gene of interest.
nucleotides that are not complementary to the region of
III. Isolation of a desired DNA fragment. template DNA
IV. Ligation of the DNA fragment into a vector. (c) the double-stranded DNA that need to be amplified
V. Insertion of recombinant DNA into the host. (d) specific sequences present on recombinant DNA
Correct order is 90 The Taq polymerase enzyme is obtained from
(a) I, II, III, IV and V (b) III, II, IV, V and I CBSE-AIPMT 2015
(c) II, IV, V, III and I (d) I, IV, V, III and II (a) Thiobacillus ferroxidans
(b) Bacillus subtilis
82 In bacterial cells, the membrane is digested with the
(c) Pseudomonas subtilis
help of enzyme
(d) Thermus aquaticus
(a) cellulase (b) lysozyme
(c) chitinase (d) lipase 91 A single PCR amplification cycle involves
(a) denaturation (b) extension
83 RNA is removed by the treatment with
(a) ribonuclease (b) protease (c) annealing (d) All of these
(c) chitinase (d) cellulase 92 The correct order of steps in Polymerase Chain
84 Proteins are removed by treatment with Reaction (PCR) is NEET 2018
(a) ribonuclease (b) chitinase (a) Denaturation, Extension, Annealing
(c) cellulase (d) protease (b) Annealing, Extension, Denaturation
(c) Extension, Denaturation, Annealing
85 DNA precipitation out of a mixture of biomolecules
(d) Denaturation, Annealing, Extension
can be achieved by treatment with NEET 2019
(a) chilled ethanol 93 The below diagram refer to PCR. Identify the steps A,
(b) methanol at room temperature B, C and D. Select the correct option.
(c) chilled chloroform Region to be amplified
(d) isopropanol
5′ 3′ dsDNA
86 Purified DNA ultimately precipitates out and this can 3′ 5′
be seen as collection of fine threads in the suspension Heat A
as seen in the figure. It refers to
5′ 3′
3′ 5′
Primers
5′ 3′
B
3′ 5′

5′ 3′
3′ 5′
C
5′ 3′
(a) DNA Spooling (b) DNA digestion 3′ 5′
(c) DNA recognition (d) DNA bands
30 cycles
87 Chimeric DNA is AIIMS 2019
(a) gene clone (b) recombinant DNA
(c) transposon (d) vector shuttle D

88 Polymerase Chain Reaction (PCR) needs

(a) DNA template (b) Primers (a) A–Denaturation at 94-96°C, B–Annealing at 40-60°C,
C–Extension through Taq polymerase at 72°C,
(c) Taq polymerase (d) All of these D–Amplified
 (b) A–Annealing at 94-96°C, B–Denaturation at 40-60°C, (a) both transformed and untransformed recipient cells will
C–Extension through Taq polymerase at 72°C, die
D–Amplification (b) both transformed and untransformed recipient cells will
(c) A–Extension through Taq polymerase at 40-60°C, grow
B–Amplification, C–Denaturation at 40-60°C, (c) transformed recipient cells will grow and untransformed
D–Annealing at 94-96°C
recipient cells will die
(d) A–Amplification, B–Extension through Taq polymerase
at 40-60°C, C–Denaturation at 40-60°C, D–Annealing (d) transformed recipient cells will die and untransformed
at 94-96°C recipient cells will grow

94 If a recombinant DNA bearing gene for ampicllin 95 Protein encoding gene which is expressed in
resitance is transferred into E. coli and the host cells heterologous host is
are spread on agar plates containing ampicillin, then (a) foreign protein (b) heterologous protein
AIIMS 2018 (c) recombinant protein (d) alien protein

TOPIC 4~ Bioreactors and Downstream Processing

96 In continuous culture system,
(a) used medium is drained out A B C D E
(b) biomass produced is high (a) Motor Foam Sterile air Steam for Acid/base
(c) no new medium is added breaker sterilisation for pH
(d) Both (a) and (b) control

97 A bioreactor (b) Foam Sterile air Steam for Acid/Base Motor

breaker sterilisation for pH
(a) is hybridoma control
(b) cultures products containing radioactive isotopes
(c) Acid/base Motor Foam Sterile air Steam for
(c) cultures for the synthesis of new chemicals for pH breaker sterilisation
(d) cultures large volume of living cells control
98 Stirred-tank bioreactors have been designed for the (d) Sterile air Steam for Foam Motor Acid/base
(a) purification of the product sterilisation breaker for pH
control
(b) addition of preservatives to the product
(c) availability of oxygen throughout the biorector 101 Identify the correct match for the given apparatus.
(d) ensuring anaerobic conditions in the culture vessel
99 Stirred-tank bioreactors are advantageous over shake
flasks because they
(a) provide high temperature and pH
(b) provide better aeration and mixing properties
(c) do not allow the entry of CO2
(d) are easy to operate
100 Simple stirred-tank bioreactor is given below. Identify
A, B, C, D and E.

A
E
B

Apparatus Functions
D Flat bladed impeller
(a) Gene gun Vectorless direct gene transfer
Culture broth (b) Column chromatograph Separation of chlorophyll
pigments
(c) Sparged tank bioreactor Carry out fermentation process
(d) Respirometer Finding out rate of respiration
C
102 The components of a bioreactor are 103 The process of separation and purification of
I. an agitator system. expressed protein before marketing is called
II. an oxygen delivery system. (a) upstream processing NEET 2017
III. foam control system. (b) downstream processing
IV. temperature control system. (c) bioprocessing
V. pH control system. (d) post-production processing
VI. sampling ports to withdraw cultures periodically. 104 Which of the following is not a component of
Choose the correct option. downstream processing? NEET 2016
(a) I, II, III, IV and V (b) II, IV, V and VI (a) Separation (b) Purification
(c) I, II, III, IV and VI (d) All of these (c) Preservation (d) Expression

NEET
SPECIAL TYPES QUESTIONS
I. Assertion and Reason 110 Assertion (A) Restriction endonucleases are also
■ Direction (Q. No. 105-113) In each of the following called ‘molecular scissors’.
questions, a statement of Assertion (A) is given followed Reason (R) When fragments generated by restriction
by corresponding statement of Reason (R). Of the endonucleases are mixed, they join together due to
statements, mark the correct answer as their sticky ends.
(a) If both A and R are true and R is the correct explanation 111 Assertion (A) All endonucleases cut DNA at specific
of A sites.
(b) If both A and R are true, but R is not the correct
explanation of A Reason (R) Endonucleases were discovered from
(c) If A is true, but R is false viruses.
(d) If A is false, but R is true 112 Assertion (A) The tumour inducing plasmid
(Ti plasmid) of Agrobacterium tumefaciens acts as a
105 Assertion (A) Biotechnology deals with techniques
cloning vector in recombinant DNA technology.
that use living organism to produce products useful
for humans. Reason (R) The Ti plasmid which is used in the
mechanisms of delivering genes to a cell remains
Reason (R) It uses only a unicellular organism.
pathogenic.
106 Assertion (A) Maintenance of sterile environment is
113 Assertion (A) Use of chitinase enzyme is necessary
essential for manufacture of biotechnological
for isolation of DNA from fungal cells.
products.
Reason (R) Fungal cell wall is made up of chitin and
Reason (R) This is to enable growth of desired chitinase is able to digest it.
prokaryotic or eukaryotic cells.
107 Assertion (A) Origin of replication is an essential II. Statement Based Questions
part of a vector.
114 Which one is a true statement regarding DNA
Reason (R) Ori is responsible for initiating
polymerase used in PCR? CBSE-AIPMT 2012
replication.
(a) It is used to ligate introduced DNA in recipient cells
108 Assertion (A) Foreign DNA and vector DNA cut (b) It serves as a selectable marker
with the help of ligase. (c) It is isolated from a virus
Reason (R) Ligase acts by forming phosphodiester (d) It remains active at high temperature
bonds. 115 Following statements describe the characteristics of
109 Assertion (A) In gel electrophoresis, DNA fragments the enzyme restriction endonulease. Identify the
are separated. incorrect statement. NEET 2019
Reason (R) DNA is negatively charged, so it moves (a) The enzyme binds DNA at specific sites and cuts only
towards anode under electric field. one of the two strands
 (b) The enzyme cuts the sugar-phosphate backbone at (c) Copy number refers to the number of copies of plasmid
specific sites on each strand present in a cell
(c) The enzyme recognises a specific palindromic (d) Copy number of plasmid varies from 50-100 per cell
nucleotide sequence in the DNA
122 Which of the statements given is incorrect?
(d) The enzyme cuts DNA molecules at identified position
(a) In microinjection method, foreign DNA is directly
within the DNA
injected into the nucleus of animal cell by using
116 Which of the following statement is incorrect? microneedles
(a) Nucleic acid is fragmented by nucleases (b) Microinjection method is used in oocytes, eggs and
(b) Construction of recombinant DNA involves cleaning embryo
DNA segments with endonuclease and rejoining with (c) Electroporation is the formation of temporary pores
ligase in the plasma membrane of host cell by using
(c) Genetic engineering is making artificial limbs and lysozyme or calcium chloride
diagnostic instruments (d) In chemical mediated gene transfer method, certain
(d) Ti plasmid transforms cells of plants chemicals such as Ca phosphate help foreign DNA to
enter the host cell
117 Which of the following statement is incorrect?
(a) DNA being a hydrophilic molecule cannot pass through 123 Consider the following statements.
cell membranes I. Recombinant DNA technology popularly known as
(b) Agrobacterium tumefaciens delivers a piece of DNA genetic engineering is a stream of biotechnology
known as ‘Z-DNA’ which transforms normal plant cells which deals with the manipulation of genetic material
into tumour cells and directs these tumour cells to by man in vitro.
produce chemicals against pathogens
II. pBR322 is the first artificial cloning vector developed
(c) Retrovirus, adenovirus, papillomavirus are also now
used as cloning vectors in animal because of their in 1977 by Boliver and Rodriquez from E. coli
ability to transform normal cells into cancerous cell plasmid.
(d) In genetic engineering, DNA from different sources are III. Restriction enzymes belong to a class of enzymes
cut with the same restriction enzymes so that both DNA called nucleases.
fragments have same kind of sticky ends
Which of the statements given above are correct?
118 Consider the following statements and select the (a) I and II (b) I and III (c) II and III (d) I, II and III
correct option.
124 Given below are four statements pertaining to
(a) A soil inhabiting plant bacterium, Agrobacterium
tumefaciens, a pathogen of several dicot plants is able to
separation of DNA fragments using gel
transfer a piece of DNA known as T-DNA electrophoresis. Identify the incorrect statements.
(b) The T-DNA causes tumours NEET (Odisha) 2019
(c) Tumour formation is induced by Ti plasmid I. DNA is negatively charged molecule and so it is loaded
(d) All of the above on gel towards the anode terminal.
II. DNA fragments travel along the surface of the gel
119 Which of the following statement is incorrect? whose concentration does not affect movement of
(a) Each restriction endonuclease recognises a specific DNA.
palindromic nucleotide sequence
III. Smaller the size of DNA fragment larger is the distance
(b) Specific base sequence is known as recognition
sequence it travels through it.
(c) Restriction enzymes cannot cut DNA IV. Pure DNA can be visualised directly by exposing
(d) Restriction enzymes belong to enzymes called UV-radiation.
nucleases Select the correct option from the following.
120 Which of the following statement is incorrect? (a) I, III and IV (b) I, II and III
(a) EcoRI cuts the DNA between bases G and A (c) II, III and IV (d) I, II and IV
(b) Making multiple identical copies of any template DNA 125 Study the given figure carefully and select the correct
is called cloning statements regarding this.
(c) pBR322 is a natural plasmid Wells
(d) Agrobacterium tumefaciens is a natural genetic engineer DNA
bands
121 Choose the incorrect statement . A B
(a) Ori also controls the copy numbers of the linked DNA
(b) If a foreign DNA ligates at the Bam HI site of
tetracycline resistance gene in the vector pBR322, the
recombinant plasmid loses the tetracycline resistance
due to insertion of foreign DNA
 I. It represents typical agarose gel electrophoresis 130 Consider the following statements.
showing differential migration of DNA fragments. I. Bioreactors are vessels of large volumes in which raw
II. Lane 1 contains undigested DNA fragments. materials are biologically converted into specific
III. Lanes 2 to 4 contain digested DNA fragment. products.
IV. Smallest DNA bands are present at A position and II. One of the most commonly used bioreactor is of
largest DNA bands are present at B position. stirring type.
(a) I, II and III (b) I, II and IV III. Shake flasks are used for growing and mixing the
(c) II and III (d) III and IV desired materials on a small scale in the laboratory.
IV. A large scale production of desired biotechnological
126 Read the statements about gene gun method. product is done by using ‘bioreactors’.
I. This method is also known as biolistic technique. Which of the statements given above are correct?
II. In this method, cells are bombarded with high velocity (a) I and II (b) I and III
microparticles of gold or tungsten coated with DNA in (c) I, II and III (d) I, II, III and IV
plants.
131 Which statement is correct?
III. Important crop plants like maize, rice and wheat have
I. The downstream processing and quality control testing
now been transformed by this method.
vary from product to product.
Which of the statements given above are correct? II. In bioreactors, raw materials are biologically
(a) I and II (b) I and III (c) II and III (d) I, II and III converted into specific products.
127 Identify the correct statements. III. Large amount of recombinant protein can be produced
I. The first recombinant DNA was constructed by using a by gene cloning.
piece of DNA from plasmid carrying antibiotic- IV. pBR322 vector was constructed by using DNA
resistance gene in the bacterium Salmonella derived from naturally occurring plasmids of E. coli.
typhimurium and linked it to the plasmid of E. coli. (a) I, II and III (b) Only IV
II. When cut by the same restriction enzyme, the resultant (c) II, III and IV (d) All of these
DNA fragments have the same kind of sticky ends and
these can be joined together using DNA ligases. 132 Which statement is incorrect ?
III. The presence of more than one recognition sites within I. Retroviruses have also been disarmed and are now
the vector will generate several fragments, which will used to deliver desirable genes into animals cells.
complicate the gene cloning. II. Downstream processing is one of the steps of R-DNA
(a) I, II and III (b) I and II technology.
(c) Only I (d) II and III III. DNA is a negatively charged molecule.
128 Read the statements. IV. The presence of chromogenic substrate gives blue
I. In the process of recombinant DNA technology, after colour colonies, if the plasmid in the bacteria does not
several treatment the purified DNA is precipitated by have an insert.
adding chilled acetone. (a) I and II (b) I, III and IV
(c) All of these (d) None of these
II. The bacterial/plant, animal cell is broken down by
enzymes to release DNA, along with RNA, proteins, 133 For selectable marker.
polysaccharides and lipids. I. It helps to select the host cells which contain the vector
Choose the correct option for above statements. and eliminate the non-transformants.
(a) I is true, but II is false (b) I is false, but II is true II. Genes encoding resistance to antibiotics like ampicillin,
(c) I and II are true (d) I and II are false chloramphenicol, tetracycline or kanamycin, are useful
selectable markers for E.coli.
129 Which of the following statements are correct with
respect to a bioreactor? Which of the statements given above are correct?
(a) Only I (b) Only II
I. It can process small volume of culture.
(c) I and II (d) None of these
II. It provides optimum temperature, pH, salt, vitamins
and oxygen. 134 I. DNA being a hydrophilic molecule cannot pass
through cell membranes.
III. Sparged stirred-tank bioreactor is a stirred type reactor
II. The bacteria should be made competent to accept the
in which air is bubbled.
DNA molecule.
Choose the correct option.
The correct option regarding the above statements is
(a) I and II (b) I and III
(c) II and III (d) I, II and III (a) I is true, but II is false (b) II is true, but I is false
(c) I and II are true (d) I and II are false
III. Matching Type Questions 139 Match the following columns.
Column I Column II
135 Match the following columns.
A. PCR 1. Join or hybridise
Column I Column II
B. Denaturation of DNA 2. Polymerisation
A. Recombinant DNA 1. Sea weeds
C. Annealing 3. Thermus aquaticus
B. Gel electrophoresis 2. DNA staining
D. Extension 4. Kary Mullis
C. Ethidium bromide 3. Plasmid DNA that has
incorporated human DNA Codes
D. Agarose 4. Process by which DNA fragments
A B C D A B C D
are separated based on their size (a) 4 3 2 1 (b) 1 2 3 4
(c) 3 1 2 4 (d) 4 3 1 2
Codes
140 Match the following columns.
A B C D A B C D
(a) 3 4 2 1 (b) 3 2 1 4 Column I Column II
(c) 2 1 4 3 (d) 3 4 1 2 A. Plasmids 1. Natural polymer from sea water
136 Match the Column I with Column II with respect to B. Bacteriophages 2. Hybrid vector derived from plasmids
the nomenclature of enzyme Eco RI and select the
C. Cosmids 3. Virus infecting bacteria
correct answer from codes given below.
D. Agarose 4. Circular, extrachromosomal DNA
Column I Column II
A. E 1. Ist in order of identification Codes
A B C D A B C D
B. co 2. genus
(a) 2 1 3 4 (b) 4 3 2 1
C. R 3. species (c) 3 2 1 4 (d) 1 4 3 2
D. I 4. strain 141 Match the following columns.
Codes Column I Column II
A B C D A B C D (Scientists) (Contributions)
(a) 3 4 1 2 (b) 2 3 4 1 A. Arber, Nathan and 1. Term biotechnology
(c) 2 1 4 3 (d) 2 3 1 4 Hamilton Smith

137 Match the following columns. B. Paul Berg 2. First recombinant DNA
C. Herbert Boyer and 3. Father of genetic engineering
Column I Column II Stanley Cohen
A. Bacterial cell is treated with 1. Lysozyme D. Karl Erkey 4. Isolated first restriction
B. Plant cell is treated with 2. Cellulase endonuclease from bacteria

C. Fungal cell is treated with 3. Chitinase Codes

A B C D A B C D
Codes (a) 1 4 3 2 (b) 3 2 1 4
A B C A B C (c) 4 3 2 1 (d) 4 3 1 2
(a) 3 2 1 (b) 2 3 1
142 Match the following columns.
(c) 1 2 3 (d) 3 1 2
138 Match the following columns. Column I Column II
(Vectors) (Derivative microorganisms)
Column I Column II A. Eco RI 1. E. coli R 245
A. Isolation of purified DNA 1. Restriction enzyme B. Hind III 2. Bacillus amyloliquefaciens
B. Cutting of DNA at specific 2. Gel electrophoresis C. Bam HI 3. Haemophilus influenzae
location
D. Eco RII 4. Escherichia coli RY13
C. Isolation of DNA fragments 3. Chilled ethanol
D. Amplification of gene 4. PCR Codes
A B C D
Codes (a) 1 2 3 4
A B C D A B C D (b) 3 2 1 4
(a) 1 2 3 4 (b) 3 1 2 4 (c) 4 3 2 1
(c) 2 1 3 4 (d) 3 2 1 4 (d) 4 2 3 1
CHAPTER 11 > Biotechnology : Principles and Processes

143 Match the following enzymes with their functions. 144 Match the following columns and choose the correct
NEET (Odisha) 2019 option from the codes given below. AIIMS 2019

Column I Column II Column I Column II

A. Restriction 1. Joins the DNA fragments (Substrates) (Enzymes)
endonuclease A. Ribonucleotide 1. Chitinase
B. Restriction 2. Extends primers on genomic DNA
B. Chitin 2. Cellulase
exonuclease template
C. DNA ligase 3. Cuts DNA at specific position C. Cellulose 3. Ribonuclease

D. Taq polymerase 4. Removes nucleotides from the ends of

Codes
DNA
A B C
Codes (a) 1 2 3
A B C D A B C D (b) 3 1 2
(a) 3 1 4 2 (b) 3 4 1 2 (c) 3 2 1
(c) 4 3 1 2 (d) 2 4 1 3 (d) 2 1 3

NCERT Exemplar
MULTIPLE CHOICE QUESTIONS
145 A bacterial cell was transformed with a recombinant 150 Which of the following is not required in the
DNA molecule that was generated using a human preparation of a recombinant DNA molecule?
gene. However, the transformed cells did not produce (a) Restriction endonuclease (b) DNA ligase
the desired protein. Reasons could be (c) DNA fragments (d) E. coli
(a) human gene may have intron which bacteria cannot 151 The role of DNA ligase in the construction of a
process
recombinant DNA molecule is
(b) amino acid codons for humans and bacteria are different
(c) human protein is formed but degraded by bacteria (a) formation of phosphodiester bond between two DNA
(d) All of the above fragments
(b) formation of hydrogen bonds between sticky ends of
146 ‘Restriction’ in restriction enzyme refers to
DNA fragments
(a) cleaving of phosphodiester bond in DNA by the enzyme
(c) ligation of all purine and pyrimidine bases
(b) cutting of DNA at specific position only
(d) None of the above
(c) prevention of the multiplication of bacteriophage by the
host bacteria 152 In agarose gel electrophoresis, DNA molecules are
(d) All of the above separated on the basis of their
147 Which of the following bacteria is not a source of (a) charge only (b) size only
restriction endonuclease? (c) charge to size ratio (d) All of these
(a) Haemophilus influenzae 153 Which of the given statement is correct in the context
(b) Escherichia coli of visualising DNA molecules separated by agarose
(c) Agrobacterium tumefaciens gel electrophoresis?
(d) Bacillus amyloliquefaciens (a) DNA can be seen in visible light
148 Which of the following enzymes catalyses the (b) DNA can be seen without staining in visible light
removal of nucleotides from the ends of DNA? (c) Ethidium bromide stained DNA can be seen in visible
(a) endonuclease (b) exonuclease light
(c) DNA ligase (d) Hind II (d) Ethidium bromide stained DNA can be seen under
149 Which of the following statements does not hold true exposure to UV-light
for restriction enzyme? 154 The most important feature in a plasmid to serve as a
(a) It recognises a palindromic nucleotide sequence vector in gene cloning experiment is
(b) It is an endonuclease (a) origin of replication (ori)
(c) It is isolated from viruses (b) presence of a selectable marker
(d) It can produce the same kind of sticky ends in different (c) presence of sites for restriction endonuclease
DNA molecules (d) its size
155 An antibiotic resistance gene in a vector usually helps in 159 Which of the following contributed popularising
the selection of the PCR (Polymerase Chain Reaction)
(a) competent bacterial cells technique?
(b) transformed bacterial cells (a) Easy availability of DNA template
(c) recombinant bacterial cells (b) Availability of synthetic primers
(d) None of the above (c) Availability of cheap deoxyribonucleotides
156 The transfer of genetic material from one bacterium to (d) Availability of ‘thermostable’ DNA polymerase
another through the mediation of a viral vector is termed as 160 Who among the following was awarded the
(a) transduction Nobel Prize for the development of PCR
(b) conjugation technique?
(c) transformation (a) Herbert Boyer (b) Har Govind Khorana
(d) translation (c) Kary Mullis (d) Arthur Kornberg
157 Significance of heat shock method in bacterial 161 Which of the following steps are catalysed by
transformation is to facilitate Taq polymerase in a PCR reaction?
(a) binding of DNA to the cell wall (a) Denaturation of template DNA
(b) uptake of DNA through membrane transport proteins (b) Annealing of primers to template DNA
(c) uptake of DNA through transient pores in the bacterial cell (c) Extension of primer end on the template DNA
wall (d) All of the above
(d) expression of antibiotic resistance gene 162 Which of the following should be chosen for best
158 While isolating DNA from bacteria, which of the yield if one were to produce a recombinant
following enzymes is not required? protein in large amounts?
(a) Lysozyme (a) Laboratory flask of largest capacity
(b) Ribonuclease (b) A stirred-tank bioreactor without in-lets and out-lets
(c) Deoxyribonuclease (c) A continuous culture system
(d) Protease (d) All of the above

Answers
‡ Mastering NCERT with MCQs
1 (c) 2 (d) 3 (a) 4 (d) 5 (a) 6 (a) 7 (b) 8 (c) 9 (b) 10 (d)
11 (a) 12 (b) 13 (d) 14 (c) 15 (b) 16 (d) 17 (c) 18 (c) 19 (a) 20 (a)
21 (c) 22 (a) 23 (c) 24 (c) 25 (c) 26 (b) 27 (b) 28 (a) 29 (b) 30 (c)
31 (b) 32 (b) 33 (b) 34 (b) 35 (d) 36 (c) 37 (a) 38 (d) 39 (b) 40 (a)
41 (c) 42 (d) 43 (a) 44 (b) 45 (b) 46 (a) 47 (d) 48 (b) 49 (b) 50 (a)
51 (b) 52 (c) 53 (b) 54 (c) 55 (c) 56 (d) 57 (a) 58 (a) 59 (d) 60 (a)
61 (d) 62 (a) 63 (a) 64 (c) 65 (a) 66 (b) 67 (b) 68 (b) 69 (a) 70 (b)
71 (b) 72 (b) 73 (c) 74 (b) 75 (b) 76 (b) 77 (d) 78 (a) 79 (d) 80 (a)
81 (b) 82 (b) 83 (a) 84 (d) 85 (a) 86 (a) 87 (b) 88 (d) 89 (a) 90 (d)
91 (d) 92 (d) 93 (a) 94 (c) 95 (c) 96 (d) 97 (d) 98 (c) 99 (b) 100 (a)
101 (c) 102 (d) 103 (b) 104 (d)

‡ NEET Special Types Questions

105 (c) 106 (a) 107 (a) 108 (d) 109 (a) 110 (b) 111 (c) 112 (c) 113 (a) 114 (d)
115 (a) 116 (c) 117 (b) 118 (d) 119 (c) 120 (c) 121 (d) 122 (c) 123 (d) 124 (d)
125 (a) 126 (d) 127 (a) 128 (b) 129 (c) 130 (d) 131 (d) 132 (d) 133 (c) 134 (c)
135 (a) 136 (b) 137 (c) 138 (b) 139 (d) 140 (b) 141 (c) 142 (c) 143 (b) 144 (b)
‡ NCERT Exemplar Questions
145 (a) 146 (b) 147 (c) 148 (b) 149 (c) 150 (d) 151 (a) 152 (b) 153 (d) 154 (a)
155 (b) 156 (a) 157 (c) 158 (c) 159 (d) 160 (c) 161 (c) 162 (c)
CHAPTER 11 > Biotechnology : Principles and Processes 279

Answers & Explanations

5 (a) The two core technologies that enabled birth of 25 (c) An enzyme catalysing the removal of nucleotides
modern biotechnology are from ends of DNA is an exonuclease.
l
Bioprocess engineering, i.e. processes that help the Other options are
growth of the desired prokaryotic or eukaryotic cell in l
Endonucleases make cuts at specific positions within
large quantities in a sterile medium for the manufacture the DNA. DNA ligase joins the DNA fragments.
and multiplication of biotechnological products. l
Proteases are protein-degrading enzymes.
l
Genetic engineering, which is the process of l
DNA ligase joins two complementary single-stranded
manipulation of genes, to introduce into the host DNA molecules.
organisms.
26 (b) Restriction endonucleases recognise a specific
6 (a) Genetic engineering includes techniques which alter DNA base sequence (recognition sequence,
the chemistry of genetic material (DNA and RNA) to recognition site), bind to the DNA and then cleave
induce the expression of genes which will give the both the strands of the DNA at those specific points.
desired products.
27 (b) Restriction endonuclease enzymes cut
10 (d) Stanley Cohen and Herbert Boyer generated the first double-stranded DNA molecules at specific sites called
recombinant DNA molecule by linking a gene encoding recognition sites which have specific base sequences.
antibiotic resistance with a native plasmid of Salmonella
typhimurium. 28 (a) Restriction endonucleases recognise their specific
sequence and bind to the DNA and cut each of the two
11 (a) The linking of the anitbiotic resistance genes with strands of the double helix at specific points in their
the plasmid vector became possible with DNA ligase. It sugar-phosphate backbone.
is the enzyme that joins two complementary nucleotides.
30 (c) Palindrome in DNA is the sequence of base pairs
12 (b) The correct sequence of step of genetic engineering which read same on the two strands when orientation of
are as follows reading is kept the same, e.g.
The first step involves the identification of DNA with
5′ – GAATTC – 3′
l

desirable genes.
3′ − CTTAAG − 5′
l
These genes are introduced into the host and are
maintained in the host. The sequence given above reads the same on both the
l
Finally, the transfer of this DNA to the host progeny strands either in 5′ → 3′ direction or in 3′ → 5′
takes place. direction.
14 (c) The enzymes, commonly used in genetic 31 (b) Restriction enzymes cut DNA strands a little away
engineering are restriction endonuclease and ligase. from the centre of the palindrome site between the same
Restriction endonuclease make cuts at specific positions two bases on the opposite strands.
within the DNA and these DNA fragments can be joined 32 (b) Same restriction enzymes are used to cut both the
together end-to-end by using ligase enzyme. foreign DNA and the vector, so that the resultant DNA
16 (d) Hind II is a restriction endonuclease. It was the first fragments have the complementary kind of sticky
restriction endonuclease to be discovered. Restriction ends.
endonucleases are the enzymes which are used for 33 (b) If a linear DNA molecule is digested using a
cutting of DNA at specific location. restriction enzyme having 4 recognition sites, it will
17 (c) The restriction enzyme responsible for the cleavage produce 5 fragments.
of the given sequence is Hind II. 34 (b) When a closed circular DNA molecule is digested
↓ with a restriction enzyme having six recognition sites, it
Answers & Explanations

5′ – GTC GAC – 3′ will produce 6 DNA fragments.

3′ − CAG CTG − 5′
35 (d) DNA ligases (genetic gum) are used in recombinant
↑
DNA technology to join two individual fragments of
20 (a) The convention for naming restriction enzymes is as double-stranded DNA by forming phosphodiester bonds
follows between them to produce a recombinant DNA
The first letter of the name comes from the genus (A), (plasmid).
the second two letters come from the species (B) and the 36 (c) The enzyme DNA ligase is also called as molecular
fourth letter comes from the strain (C) of bacteria (D), glue, as it works to repair broken DNA by joining two
e.g. Eco RI comes from Escherichia coli RY13. complementary nucleotides in a DNA strand. It is
23 (c) Restriction enzymes belong to a larger class of commonly used in genetic engineering to do the reverse
enzymes called nucleases, which are of two kinds, i.e. of restriction enzyme, i.e. to join together
exonucleases and endonucleases. complementary restriction fragments.
 Other options are 54 (c) Plasmid is not single-stranded. Plasmid is an
l
DNA gyrase is an enzyme of class topoisomerase that extrachromosomal, double-stranded, circular DNA
relieves torque or strain in DNA during DNA molecule, having the ability of self-replication. These
replication. are usually found in bacterial cells and from some
l
DNA helicase unwinds the double helix to make site of yeasts. Discovery of plasmid has led to the revolution in
DNA synthesis. the biotechnological research.
l
DNA polymerase catalyses the formation of new DNA 55 (c) Plasmid is a vector which can clone small DNA
on template strand. fragments. Small fragment of DNA (about 10 Kbp size)
37 (a) Enzyme A is Eco RI which is used in step I and
that is physically separate from and can replicate freely
of chromosomal DNA within a cell.
enzyme B is DNA ligase, used in step II. Eco RI cuts
the DNA between bases G and A only when the 57 (a) pBR322 is a plasmid vector. A plasmid is an
sequence GAATTC is present in the DNA. This extrachromosomal genetic elements of DNA that is
enzyme cuts both DNA strands at the same site. capable of replicating independently of the host
Enzyme ligase joins the complementary DNA chromosome.
fragments at their sticky ends to form the recombinant 59 (d) Option (d) contains the correctly identified
DNA. components as amp R (ampicillin resistance gene) and
42 (d) A molecule of DNA can be cut into fragments by tet R (tetracycline resistance gene) are antibiotic
the enzyme restriction endonucleases. These fragments resistance genes. Rest of the options are incorrect and
of DNA can be separated on the basis of size by a can be corrected as
technique of gel electrophoresis. It is the most common l
Ori is origin of replication,
method used to separate DNA molecules on the basis of l
rop codes for proteins involved in replication of the
their size. plasmid and stands for regulation of origin of plasmid.
43 (a) Agarose is extracted from sea weeds and is a l
Hind III as well as Eco RI are restriction sites.
polysaccharide. In gel electrophoresis, DNA fragments
separate according to their size through seiving effects 60 (a) The two antibiotic resistance genes on the cloning
provided by the agarose gel. vector pBR322 are for ampicillin and tetracycline
antibiotics. Cloning vectors are DNA molecules that
44 (b) Gel electrophoresis is used for the separation of carry a foreign DNA segment and replicate inside host
molecules of similar electric charge on the basis of their cell. Plasmid in E.coli is a cloning vector.
size. Hence, smaller size of the DNA fragment, the
farther it moves on agarose gel during gel 61 (d) Origin of replication is the sequence from where
electrophoresis. replication starts and any piece of DNA when linked
to this sequence can be made to replicate within host
45 (b) DNA being negatively charged migrates towards
cells.
the positive terminal, the anode, but is loaded at
cathode. This sequence is also responsible for controlling the
copy number of the linked DNA. Thus, the function of
46 (a) The smallest fragments of DNA are found near the
ori in a vector is to help in replication of linked DNA
positive electrodes as DNA is negatively charged.
These fragments travel towards anode (farthest away and control the copy number of the linked DNA.
from the leading wells). 62 (a) A gene, whose expression helps to identify
47 (d) The DNA fragments separated on an agarose gel transformed cells is known as selectable marker.
can be visualised after staining with ethidium bromide. Usually, the genes encoding resistance to antibiotics,
It is an intercalating, fluorescent agent. The stained such as tetracycline, ampicillin, etc., are called
DNA fragments are seen as bright orange coloured selectable markers (for E. coli).
bands under UV light. 63 (a) To facilitate cloning, a vector requires a selectable
Answers & Explanations

51 (b) In recombinant DNA technology, vector refers to a marker, which helps in identifying and eliminating
plasmid used to transfer foreign DNA into a host cell, non-transformants and selectively permitting the growth
where the genes may be amplified (gene cloning) or of the transformants.
otherwise manipulated. 64 (c) Gene encoding resistance to antibiotics like
52 (c) Bacteriophage are used in recombinant DNA ampicillin, chloramphenicol, tetracycline or
technique. These are used as vectors due to their high kanamycin, useful for cloning are called selectable
number per cell and high copy numbers of their markers. These are suitable selectable markers for
genome within the bacterial cells. E. coli as the normal E. coli cells do not carry
53 (b) During ‘gene cloning’ the plasmid vector is called resistance against any of these antibiotics.
a gene taxi. Desired genes are inserted into plasmids, 65 (a) The presence of more than one recognition sites
then the plasmid with the added gene are delivered into within the vector will generate several fragments, which
a living bacterium. will complicate the gene cloning process.
68 (b) Insertional inactivation is used to differentiate 87 (b) A chimeric DNA is a recombinant DNA generated
recombinants and non- recombinants, on the basis of the by ligating sequences derived from different sources.
inability of the recombinants to produce colour in the 88 (d) PCR is a technique of synthesising multiple copies
presence of a chromogenic substrate. of the desired gene (or DNA) of interest in vitro. The
69 (a) In insertional inactivation, the recombinant DNA is basic requirements of PCR are DNA template, two sets
inserted within the coding sequence of the enzyme of primers and the enzyme (Taq polymerase).
β-galactosidase. This results into inactivation of the 90 (d) Taq polymerase is a thermostable DNA polymerase
gene synthesising this enzyme. obtained from Thermus aquaticus. This enzyme is not
70 (b) Recombinant colonies in insertional inactivation are denatured at high temperature. Thermus aquaticus is a
differentiated from non-recombinants on the basis of bacterium that lives in hot springs and hydrothermal
their inability to produce the blue colour in the presence vents at a temperature of 300ºC due to the presence of
of a chromogenic substrate. The presence of a this enzyme.
chromogenic substrate gives blue coloured colonies, if 94 (c) Transformation is a procedure through which a piece
the plasmid in bacteria does not have an insert. The of DNA is introduced in a host bacterium. Cells
presence of insert results in the inactivation of enzyme containing DNA with ampicillin resistance gene
β-galactosidase due to which colourless recombinant (transformed recipient cells) when introduced into agar
colonies are produced. plates containing ampicillin will grow, while cells
71 (b) Agrobacterium tumefaciens, a pathogen of several containing DNA without ampicillin resistance gene
dicot plants is able to deliver a piece of DNA known (untransformed recipient cells) will die. The genes
as T-DNA to transform normal plant cells into encoding resistance to antibiotics such as ampicillin,
tumour cells. chloramphenicol, tetracycline or kanamycin, etc., are
74 (b) In Ti plasmid, the virulent gene is removed to make hence, considered as useful selectable markers for
it a suitable (competent) vector for gene transfer. In it, E.coli. The normal E. coli cells do not carry resistance
tumour forming virulent genes are replaced with desired against any of these antibiotics.
genes and this way it acts as a vector to carry the 96 (d) In continuous culture system, the used medium is
desired gene to the host organism where it gets continuously drained out from one side, while
expressed to produce the desired product. simultaneously fresh medium is added from the other to
maintain the cells in their physiologically most active
76 (b) Host is made competent by treating them with
log phase. This type of method produces a larger
specific concentration of a divalent cation such as biomass leading to higher yield of desired products.
calcium, which increases the efficiency with which
DNA enters the bacterium through the creation of 99 (b) A stirred-tank bioreactor is more advantageous than
shake flasks as these provide better aeration and mixing
pores in its cell wall or cell membrane.
properties. It has an agitator system to mix the contents
79 (d) Biolistics or gene gun method is a direct or vector properly and also an oxygen delivery and also system to
less way used to introduce alien DNA into host cells. In ensure better availability of oxygen.
this method of gene transfer, high velocity micro
103 (b) The process of separation and purification of
particles of gold or tungsten, coated with DNA are
expressed proteins before marketing is called as
bombarded on the desired host cells which are usually
downstream processing. In this process, a whole range
plant cells.
of biochemical separation and purification techniques
80 (a) DNA transfer with high velocity microparticles is are used such as drying, chromatography, solvent
present in biolistics. This technique involves extraction and distillation. After purification, several
bombardment of microscopic gold particles coated with quality control testings are done so as to make the
the foreign DNA at the cells using a compressed air product suitable for marketing.
gun. It is designed to overcome the high impermeability
104 (d) Expression is not a component of downstream
Answers & Explanations

of cell wall in plants. These particles can penetrate the

processing. Infact, processing, separation and
cell wall, the cell and the nuclear membrane to deliver
purification are collectively referred to as downstream
the DNA to the nucleus.
processing and the product has to be formulated with
82 (b) In bacterial cells, the cell wall (membrane) is suitable preservatives. The product has to be subjected
digested with the help of the enzyme lysozyme. On the through this complete series before it is ready for
other hand, marketing as a finished product.
l
Cellulase and chitinase enzymes are used for the
105 (c) Assertion is true, but Reason is false. Reason can be
digestion of plant and fungal cell walls, respectively.
corrected as
l
Lipids can be removed with the treatment of the
Biotechnology uses unicellular as well as multicellular
enzyme lipase.
organisms.
85 (a) Chilled ethanol is used to precipitate DNA out of a Biotechnology deals with techniques that use living
mixture of biomolecules. Low temperature protects the organisms to produce products useful for humans.
DNA by slowing down the activity of enzymes that
could break it apart and ethanol helps in the quick 106 (a) Both Assertion and Reason are true and Reason is
precipitation of DNA. the correct explanation of Assertion.
 Maintenance of sterile (microbial contamination-free) 115 (a) The statement in option (a) is incorrect. It can be
environment is essential in biotechnological processes to corrected as
enable growth of only the desired prokaryotic or Restriction endonuclease enzymes cut both the strands
eukaryotic cells in large quantities for manufacture of of DNA helix at specific sites in their sugar-phosphate
biotechnological products like antibiotics, vaccines, backbone. The sequences being recognised by
enzymes, etc. restriction enzymes are called palindromic sequences
107 (a) Both Assertion and Reason are true and Reason is the which have same reading frame in both 5′ → 3′ and
correct explanation of Assertion. 3′→ 5′ directions.
Origin of replication (ori) is the sequence from where Rest of the statements are correct.
replication starts and any piece of DNA when linked to 116 (c) The statements in option (c) is incorrect and can
this sequence can be made to replicate within the host be corrected as
cell. Thus, it is an essential part of a vector.
Genetic engineering helps in making recombinant
108 (d) Assertion is false, but Reason is true. Assertion can be DNA and not artificial limbs and diagnostic
corrected as instruments.
In the formation of rDNA, restriction endonucleases cut Rest of the statements are correct.
both foreign DNA and vector DNA.
117 (b) The statement in option (b) is incorrect and can be
Whereas, ligases join two DNA strands by forming corrected as
phosphodiester bonds between adjacent nucleotides.
Agrobacterium tumefaciens delivers a piece of DNA
109 (a) Both Assertion and Reason are true and Reason is the known as ‘T-DNA’, which transforms normal plant
correct explanation of Assertion. cells into tumour cells and directs these tumour cells
DNA fragments can be isolated with the help of gel to produce the chemicals required by the pathogen.
electrophoresis. In this technique, negatively charged
Rest of the statements are true.
DNA moves towards the anode (positively charged) under
an electric field through a matrix. 119 (c) The statement in option (c) is incorrect and can be
110 (b) Both Assertion and Reason are true, but Reason is not corrected as
the correct explanation of Assertion. Restriction enzymes belong to a larger class of
Restriction endonucleases are molecular scissors, as these enzymes called nuclease and can cut DNA a little
enzymes cut a DNA molecule within certain specific sites away from the center of the palindromic sites, but
called restriction sites. between the same two bases on the opposite strands.
When these fragments with sticky ends are mixed, they Rest of the statements are correct.
can be joined together (end to-end) using the enzyme, 120 (c) The statement in option (c) is incorrect and can be
DNA ligase. corrected as
111 (c) Assertion is true, but Reason is false. Reason can be pBR322 was the first artificial ideal plasmid (vector)
corrected as constructed by Boliver and Rodriguez.
Rest of the statements are correct.
Restriction endonucleases were discovered from bacteria,
not from viruses. 121 (d) The statement in option (d) is incorrect and can be
All endonucleases cut DNA at specific sites within the corrected as
DNA sequence. Some plasmids may have only one or two copies per
cell whereas others may have 15-100 copies per cell.
112 (c) Assertion is true, but Reason is false. Reason can be
corrected as Rest of the statements are correct.
The tumour inducing, Ti plasmid of Agrobacterium 122 (c) Statements in option (c) is incorrect and can be
tumefaciens has been modified into a cloning vector corrected as
Electroporation involves an electric pulse of high
Answers & Explanations

which is not pathogenic to the plants. However, it can still

used to deliver genes of interest into target plant cells. voltage applied to protoplasts/cells/tissues to make
transient (temporary) pores in the plasma membrane
113 (a) Both Assertion and Reason are true and Reason is the
which facilitates the uptake of foreign DNA.
correct explanation of Assertion.
Rest of the statements are correct.
Fungal cell wall is made up of chitin. Thus, enzyme
chitinase is required for the digestion of fungal cell wall 124 (d) Statements I, II and IV are incorrect and can be
and isolation of DNA from the cell. corrected as
114 (d) The statement in option (d) is correct about DNA
l
DNA fragments are negatively charged molecules
polymerase used in PCR. Rest of the statements are and are loaded in the middle of the gel. The
incorrect and can be corrected as fragments can be separated by forcing them to move
l
It is used to catalyse DNA synthesis. towards the anode under an electric field through a
medium/matrix.
l
It does not serve as a selectable marker. l
The concentration of gel affects the resolution of
l
It is isolated from a bacterium, Thermus aquaticus.
DNA separation.
 l
The separated DNA fragments can be visualised only 153 (d) The separated DNA fragments (by the process of gel
after staining the DNA with a compound known as electrophoresis) are visualised after staining the DNA
ethidium bromide followed by exposure to fragments with ethidium bromide followed by exposure
UV-radiation. to UV-radiation. These fragments are seen as orange
125 (a) Statements I, II and III are correct, while statement coloured bands (light).
IV is incorrect and can be corrected as 154 (a) All of the given features of a vector are important to
Largest DNA bands will be at (A) and the smallest facilitate cloning, but out of them origin of replication
DNA bands will be at (B) because in this technique, (ori) is the most important one.
DNA moves according to their size through sieving This is due to the following reasons
effect provided by the agarose gel. Hence, the smaller l
ori is a DNA sequence that is responsible for initiating
the fragment size, the farther it moves. replication. Any piece of DNA when linked to this
128 (b) Statement I is false, but II is true. Statement I can sequence can replicate within the host cells.
be corrected as l
ori also controls the copy numbers of the linked DNA.
In the process of recombinant DNA technology, after 155 (b) An antibiotic resistance gene in a vector acts as a
several treatment the purified DNA is precipitated by selectable marker and usually helps in the selection of
adding chilled ethanol.
transformed bacterial cells.
129 (c) Statements II and III are correct. Statement I is Selectable markers in a vector help in identifying and
incorrect and can be corrected as eliminating non-transformants and selectively permitting
Large volumes (100-1000 L) of culture can be the growth of the transformants.
processed in bioreactors. A bioreactor provides the
156 (a) Transduction is the process by which genetic material
optimal conditions for obtaining the desired product.
is transferred from one bacterium to another through the
In the sparged stirred-tank bioreactor, sterile air
mediation of viral vectors.
bubbles are sparged. This increases the surface area
for oxygen transfer. 157 (c) In heat shock method, firstly the cells with
145 (a) Inducing a cloned eukaryotic (human) gene to recombinant DNA are placed on ice, followed by
function in a prokaryotic (bacteria) host can be exposing them briefly to a temperature of 42°C (heat
difficult sometimes. shock) and then putting them back on ice. This enables
uptake of DNA through transient pores created in the
The presence of long non-coding introns in eukaryotic
genes may prevent the correct expression of these bacterial cell wall.
genes in prokaryotes as they lack the RNA-splicing 159 (d) Polymerase Chain Reaction (PCR) involves
machinery. amplification of specific DNA sequences, carried out
147 (c) Agrobacterium tumefaciens is not a source of in vitro.
restriction endonuclease but serves as the source of Ti Such repeated amplification is achieved by the use of a
plasmid. thermostable DNA polymerase (isolated from a bacterium
On the other hand, Hind III is obtained from (Thermus aquaticus), which remains active and stable
Haemophilius influenzae, Eco RI obtained from during the high temperature and induced denaturation of
Escherichia coli and Bam HI obtained from Bacillus double-stranded DNA. Thus, availability of thermostable
amyloliquefaciens. DNA polymerase has popularised PCR.
149 (c) The statement in option (c) is not true for 160 (c) PCR (Polymerase Chain Reaction) technique was
restriction enzymes. It can be corrected as developed by Kary Mullis in 1985 and for this he
It is isolated from bacteria and not from viruses. received Nobel Prize for chemistry in 1993.
Rest other statements are correct. Other options are
l
HG Khorana discovered DNA ligase enzyme into phage
150 (d) E. coli is not required in the preparation of
in 1969.
recombinant DNA molecules.
l
DNA polymerase was discovered by Arthur Kornberg.
On the other hand, restriction endonuclease and DNA
ligase can be used to make a stable recombinant DNA
l
Herbert Boyer generated the first recombinant DNA
molecule with DNA fragments that have been molecule by combining a gene from a bacterium with
obtained from different organisms. plasmid of E. coli in 1972.