MODERN INSTITUTE OF EDUCATION

RAJATALAB, VARANASI, 8318013115,9555590189 PRE-FOUNDATION

&FOUNDATION CLASSES 6th to 12th

Test / Exam Name: Test series Standard: 12TH SCIENCE Subject: BIOLOGY
Student Name: Section: Roll No.:

Questions: 15 Time: 105 Mins Marks: 65

Q1. Illustrate the design of a bioreactor. Highlight the difference between a flask in your laboratory and a 6 Marks
bioreactor which allows cells to grow in a continuous culture system.
Ans:
Small volume cultures cannot yield appreciable quantities of products. To produce in large quantities, the
development of bioreactors, where large volumes (100-1000 litres) of culture can be processed, was required. Thus,
bioreactors can be thought of as vessels in which raw materials are biologically converted into specific products,
individual enzymes, etc., using microbial plant, animal or human cells. A bioreactor provides the optimal conditions
for achieving the desired product by providing optimum growth conditions (temperature, pH, substrate, salts,
vitamins, oxygen).

Structure of Bioreactor:

It is a cylindrical structure with a curved base.

A stirrer is present for even mixing and oxygen availability throughout the reactor.
There is an agitator system, an oxygen delivery system, a foam control system, a temperature control system,
etc.
There is a sampling port through which small volumes of culture can be taken out periodically.

A flask in a laboratory cannot be used for producing recombinant DNA on large scale. Unlike a bioreactor; a flask
cannot be used to grow culture continuously.
Difference:
Flask Bioreactor
i. Flask is used to small i. Bioreactor is used for commercial scale production.
laboratory scale testing of a
culture.
ii. The cells harbouring cloned ii. The cells can also be multiplied in a continuous culture system wherein the used
genes of interest may be medium is drained out from one side while fresh medium is added from the other
grown on a small scale in to maintain the cells in their physiologically most active log/ exponential phase.
the laboratory. This type of culturing method produces a larger biomass leading to higher yields
of desired protein.
iii.Small volume cultures iii.To produce in large quantities, the development of bioreactors, where large
cannot yield appreciable volumes (100–1000 litres) of culture can be processed, was required.
quantities of products.
Q2. Cloning of genes, play a very significant role in genetic engineering, helping the transfer of desirable 5 Marks
foreign genes into different hosts. The scientists, to make this process easier and effective are creating
engineered vectors in such a way that they help easy linking of foreign DNA and selection of
recombinants from non recombinants. 'pBR322' is one such engineered vectors developed by scientists.
A diagram of an engineered vector pBR322 is given below:

i. Name the host for this cloning vector.

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 ii. Identify 'Rop' and 'Ori' in the diagram from 'U', 'V', 'W', 'X', 'Y' and 'Z'. Write their functions.
iii. Draw the fragments that will be formed by the action of 'Z' (marked in the diagram) on the specific
site of the DNA segment given below:
5'-GTACGAATTCCTGA-3'
3'-CATGCTTAAGGACT-5'
Ans:
a. E.coli/ Escherichia coli.
b.

rop—‘W’,— code for the proteins involved in the replication of the plasmid.
ori—‘U’,— this is a sequence from where replication starts/ control the copy number of the linked DNA.

iii.
5’-GTACG 3’ 5’ AATTCCTGA-3’
3’-CATGCTTAA 5’ 3’ GGACT-5’
Q3. a. Explain the different steps carried out in Polymerase Chain Reaction, and the specific roles of the 5 Marks
enzymes used.
b. Mention application of PCR in the field of:
i. Biotechnology.
ii. Diagnostics.
Ans:
a.

Enzyme DNA polymerase/ Taq polymerase, the enzyme extends the primers using nucleotide provided in the
reaction.
b.
i. Multiple copy of gene of interest can be made in vitro/ gene amplification.
ii. Early detection of disease at a time when the symptoms are not yet visible/ or the toxin is in low
concentration/ used to detect mutations in genes in suspected cancer patients/ a powerful technique to
identify many other genetic disorders.
Q4. a. Name the type of DNA that forms the basis of DNA fingerprinting and mention two features of 5 Marks
this DNA.
b. Write the steps carried out in the process of DNA fingerprinting technique, and mention its
application.
Ans:
a. Satellite DNA/ repetitive DNA.
These sequences normally do not code for any proteins, these sequence show high degree of polymorphism.
b.
i. Isolation of DNA.
ii. Digestion of DNA by restriction endonucleases.
iii. Separation of DNA fragments by electrophoresis.
iv. Transferring (blotting) of separated DNA fragments to synthetic membranes such as nitrocellulose or nylon.
v. Hybridisation using labelled VNTR probe.
vi. Detection of hybridised DNA fragments by autoradiography.
Application Forensic science/ determining population and genetic diversities/ paternity test.

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Q5. Explain the role of lactose as an inducer in 5 Marks
a lac operon.
Ans:
Lactose/inducer binds with repressor protein, inactivates it, frees operator gene, RNA polymerase freely move over
structural genes/RNA polymerase access to the promoter, transcribing to, lac mRNA, which on translation, produce
transacetylase, permease, β galactosidase.

Q6. What essential features must be present in a cloning 5 Marks

vector?
Ans:
Most commercial cloning vectors have key features that have made their use in molecular biology so widespread.
Control of Expressions: In the case of expression vectors, the main purpose of these vehicles is the controlled
expression of a particular gene inside a convenient host organism (e.g. E. coli). Control of expression can be very
important; it is usually desirable to insert the target DNA into a site that is under the control of a particular
promoter. Some commonly used promoters are T7 promoters, lac promoters and cauliflower mosaic virus's 35s
promoter (for plant vectors).
Selectable Marker: To allow for convenient and favorable insertions, most cloning vectors have had nearly all their
restriction sites engineered out of them and a synthetic multiple cloning site (MCS) inserted that contains many
restriction sites. MCSs allow for insertions of DNA into the vector to be targeted and possibly directed in a chosen
orientation. A selectable marker, such as an antibiotic resistance is often carried by the vector to allow the selection
of positively transformed cells. All plasmids must carry a functional origin of replication (ori).
Q7. Answer the following questions: 5 Marks
Describe the temperature treatment that enhances the bacteria to take up the
rDNA.
Ans:
a. Host cells are incubated with rDNA on ice.
b. Followed by placing them briefly at 42°C.
c. Then transfer them back on ice.
This enables the host cells (bacteria) to take up the rDNA.
Q8. Read the case study given below and answer the questions that follow: 4 Marks
Since DNA is a hydrophilic molecule, it cannot pass through cell membranes. To facilitate the uptake of
DNA by bacterial cells, they must be made ‘competent’ to take up DNA. This is achieved by treating
them with a specific concentration of a divalent cation, such as calcium, which enhances the efficiency
of DNA entry through pores in the cell wall. The process involves incubating the cells with recombinant
DNA on ice, followed by a brief heat shock at 42°C, and then returning them to ice. This treatment
enables bacteria to incorporate the recombinant DNA.
For introducing DNA into animal cells, the micro-injection method involves directly injecting recombinant
DNA into the nucleus of the cell. In plant cells, the gene gun method uses high-velocity micro-particles
coated with DNA to bombard the cells. Another approach involves using ‘disarmed pathogen’ vectors
that transfer recombinant DNA into the host cell when they infect it.
1. Why is DNA unable to pass through cell membranes naturally?
2. What role does calcium play in the process of making bacterial cells competent?
3. Describe the process of heat shock used to facilitate DNA uptake by bacterial cells.
OR
3. Compare the methods used for introducing recombinant DNA into animal cells and plant cells.
Ans:
1. DNA is hydrophilic and cannot pass through the hydrophobic lipid bilayer of cell membranes naturally.
2. Calcium helps increase the efficiency of DNA uptake by bacterial cells by making them more permeable.
3. The bacterial cells, incubated with recombinant DNA on ice, are subjected to a brief heat shock at 42°C. This
sudden increase in temperature causes the cell membranes to become more permeable, allowing the DNA to
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 enter the cells. The cells are then returned to ice to stabilize the membranes.
OR
3. In animal cells, recombinant DNA is introduced via micro-injection, where DNA is directly injected into the
nucleus of the cell. In contrast, for plant cells, the gene gun method is used, where high-velocity micro-particles
coated with DNA are shot into the cells. Both methods aim to integrate recombinant DNA into the host cells, but
they use different techniques suited to the cell types.
Q9. Read the case study given below and answer the questions that follow: 4 Marks
Vijay lives in a building where a robbery occurred last night. Tragically, the thieves also murdered an old
lady who lived alone in the flat next to Vijay. The police arrived and collected samples from the crime
scene. Among these samples were blood and tissue from under the old lady's nails. The police suspected
three people: the cook, the maid, and the milkman. A few days later, the police managed to catch the
real culprit—the milkman. Vijay was curious about how the police solved the case so quickly. At the
police station, he learned that the investigation was greatly aided by using the PCR (Polymerase Chain
Reaction) technique to amplify the DNA from the samples collected from the victim's nails, which then
helped in identifying the suspect.
1. What is the purpose of using PCR in the investigation described in the case?
2. How did the amplification of DNA help in solving the crime?
3. Explain the role of DNA analysis in criminal investigations, using the case as an example.
OR
3. Describe the process of PCR and its significance in forensic science, referring to the case study.
Ans:
1. PCR was used to amplify the DNA from the blood and tissue samples found under the old lady's nails to help
identify the suspect.
2. Amplified DNA allowed the police to compare the victim's DNA with the suspects' DNA, leading to the
identification of the milkman as the culprit.
3. DNA analysis plays a crucial role in criminal investigations by providing a way to identify suspects based on
genetic material found at the crime scene. In the case, DNA from the victim's nails was amplified using PCR,
which allowed for accurate comparison with the suspects' DNA, leading to the identification of the milkman.
OR
3. PCR (Polymerase Chain Reaction) is a technique used to amplify specific DNA sequences, making them easier to
analyze. In forensic science, PCR is significant as it allows for the amplification of small amounts of DNA from
crime scenes, such as those found under the victim's nails in the case. This amplification helps in creating
sufficient DNA samples for comparison and identification of suspects.

Q10. Read the case study given below and answer the questions that follow: 4 Marks
The DNA, which is transferred from one organism into another by joining it with the vehicle DNA is
called passenger or foreign DNA. Generally, three types of passenger DNAs are used. These are
complementary DNA (cDNA), synthetic DNA (sDNA) and random DNA. Complementary DNA (cDNA) is
synthesized on RNA template (usually mRNA) with the help of reverse transcriptase. Synthetic DNA
(sDNA) is synthesized on DNA template or without a template. Random DNA are small fragments
formed by breaking a chromosome of an organism in the presence of restriction endonucleases.
1. What is passenger DNA?
2. How is complementary DNA (cDNA) synthesized?
3. Describe the synthesis of synthetic DNA (sDNA).
OR
3. Explain the process of creating random DNA fragments.
Ans:
1. Passenger DNA is the DNA that is transferred from one organism into another by joining it with vehicle DNA.
2. Complementary DNA (cDNA) is synthesized on an RNA template (usually mRNA) with the help of reverse
transcriptase.
3. Synthetic DNA (sDNA) can be synthesized either on a DNA template or without any template at all. When
synthesized without a template, the sequence is artificially created based on desired specifications.
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 OR
3. Random DNA fragments are formed by breaking a chromosome of an organism in the presence of restriction
endonucleases. These enzymes cut the DNA at specific sequences, resulting in small, random fragments.

Q11. Read the case study given below and answer the questions that follow: 4 Marks
In recombinant DNA technology, the fragments of DNA generated after cutting the DNA by restriction
enzymes are separated according to their size or length by gel electrophoresis. Gel electrophoresis is
performed in a gel matrix so that molecules of similar electric charges can be separated on the basis of
size. Most commonly used matrix in gel electrophoresis is agarose. The fragments are separated under
the influence of electric field. The separated DNA fragments can be seen only after staining the DNA
with compound known as ethidium bromide (EtBr) followed by exposure to UV radiation as bright
orange band.
1. Why is agarose gel matrix preferred for gel electrophoresis in recombinant DNA technology?
2. What is the purpose of staining DNA fragments with ethidium bromide (EtBr) during gel
electrophoresis?
3. Explain the principle behind the separation of DNA fragments during gel electrophoresis.
OR
3. Describe the role of restriction enzymes in recombinant DNA technology.
Ans:
1. Agarose gel matrix is preferred because it forms a porous network that allows DNA fragments to migrate based
on size under the influence of an electric field.
2. Ethidium bromide stains DNA fragments so that they become visible under UV light, appearing as bright orange
bands, facilitating their visualization and analysis.
3. Gel electrophoresis separates DNA fragments based on their size. When an electric current is applied, DNA
fragments move through the agarose gel matrix. Smaller fragments move faster and migrate farther, while
larger fragments move slower and stay closer to the point of origin.
OR
3. Restriction enzymes cut DNA at specific recognition sequences, generating DNA fragments of varying sizes.
These fragments can then be analyzed or combined with other DNA fragments from different sources to create
recombinant DNA molecules.

Q12. Taking examples under each category, discuss upstream and downstream 4 Marks
processing.
Ans:
Upstream processing: Biotechnological processes can be separated into upstream processes and downstream
processes. The upstream process is defined as the entire process from DNA isolation and culture expansion of the
cells until final product.
Downstream processing: After completion of the biosynthetic stage, the product has to be subjected through a
series of processes before it Is ready for marketing as a finished product. The processes include separation and
purification, which are collectively referred to as downstream processing. The product has to be formulated with
suitable preservatives. Such formulation has to undergo through clinical trials as in case of drugs. Strict quality
control testing for each product is also required. The downstream processing and quality control testing vary from
product to product.
Q13. a. Explain the significance of 'palindromic nucleotide sequence' in the formation of 3 Marks
recombinant DNA.
b. Write the use of restriction endonuclease in the above process.
Ans:
a. Palindromic nucleotide sequence is the recognition (specific) sequence present both on the vector and on a
desired/alien DNA for the action of the same(specific) restriction endonuclease to act upon.
b. Same restriction endonuclease binds to both the vector and the foreign DNA, cut each of the two strands of the
double helix at specific points in their sugar phosphate backbone of recognition sequence for restriction
endonucleases/palindromic sequence of vector and foreign DNA, to cut strand a little away from the centre of the
palindrome sites, creates overhanging stretches/sticky ends.

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Q14. Name and explain the techniques used in the separation and isolation of DNA fragments to be used in 3 Marks
recombinant DNA technology.
Ans:
Gel Electrophoresis.
DNA fragments on the agarose gel are negatively charged molecules, and they move towards the anode (The
fragments separate according to their size) The separated DNA fragments can be visualised after staining with
ethidium bromide.
Followed by exposure to UV radiation Separated fragments are extracted from the gel by elution.
Q15. In an E. coli cloning vector pBR 322, state the role of the following 3 Marks
genes:
i. Ori gene.
ii. Antibiotic resistance gene.
iii. Rop gene.
Ans:
a. Ori-gene: The sequence from where replication start/ any piece of DNA when linked to this sequence can be
made to replicate within the host cell, this sequence control the copy number of linked DNA.
b. Antibiotic resistance genes: Help in identifying and eliminating non transformant from transformant/ acts as
selectable marker/ helps in ligation of alien DNA at recognition site (present in one of the two antibiotic resistance
gene).
c. Rop: Codes for proteins, involved in the replication of plasmids.

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