UNIT 4 – 12 marks/70 marks

Type(No.) MCQ(2) Assertion- 2 3 marks 4 marks (CB)/

Reason(0/1) marks(1) (1/2) 5 marks
(1)
Marks 2 0/1 2 3/6 4/5

2 marks questions

1. The image below shows the result of plating bacteria in chromogenic medium after in
corporating the gene of interest in plasmid. The plates had blue and white colonies. A
single bacterium extracted from plate I, II, III is shown below.

On the basis of your observations (a) Identify the plate(s) which is/are white. Give a
reason. (b) Identify the plate(s) which is/are blue. Give a reason.
2. (a) Write the scientific name of the source organism of the thermostable DNA polymer
ase used in PCR.
(b) State the advantage of using thermostable DNA polmerase.
3. Use the information provided in the table given below to answer the questions that
follows.
PCR steps Description Temperature (0C)
Denaturation Double-stranded DNA is denatured and A
separated into single strands
B Primers bind to the single DNA strands 550C
Extension C 720C
(a) Fill the gaps of A, B and C.
(b) Write two important applications of PCR.
4. A recombinant vector with a gene of interest within the gene of β-galactosidase enzyme
is introduced into a bacterium. Explain the method that would help in selection of non-
recombinant colonies from recombinant colonies. Why is this method of selection referr
ed to as insertional inactivation?
5. (a) What effect do eating genetically modified foods have on your genes? (b) Are foods
derived from genetically modified crops nutritionally superior?
6. In the figure below, the process of recombinant DNA technology is depicted.
What is the role of enzyme Z in the above experiment? (b) Will the outcome of this
experiment be successful? Give your opinion along with your answer.
 7. Cloning sites are required in a vector to facilitate the action of restriction enzymes duri
ng genetic engineering process. What will happen if many recognition sites are present
within the vector?
8. In the figure given below, parts A and B are the missing sites from the cloning vector.
Study the figure and answer the questions that follows.

(a) Identify the parts A and B in the given illustration. (b) What is the term given to C
and D and explain why?
9. A plasmid DNA and a linear DNA (both of same size) have one site for a restriction endo
nuclease. When cut and separated on agarose gel-electrophoresis, plasmid shows one
DNA band, while linear DNA shows two fragments. Explain.
10. EcoRI is used to cut a segment of foreign DNA and that of the vector DNA to form recom
binant DNA. Show the following with the help of schematic diagrams. (i) The site where
EcoRI act and cut both the segments. (ii) Sticky ends formed on both the segments whe
re the two DNA segments join to form a recombinant DNA.

3 marks questions

1. A farmer grew 2 varieties of corn crop in fields A and B. He grew normal corn crops in
field A and GM corn crops in field B. He observed corn borers attacked only in field A. To
control it, spores of Bt were sprayed in field A.
(a) Name the gene in the spores responsible for the control of this pest.
(b) What effect will the spores of Bt have on the insect pest?
(c) How has the field B developed resistance against this pest?
2. Lipoprotein lipase deficiency (LPLD) is a genetic disorder in which a person has a defecti
ve gene for lipase. This leads to high triglycerides, stomach pain, fat deposits under the
skin. It may eventually affect the liver, pancreas and may also cause diabetes. The disor
der occurs if a child acquires defective genes from both parents (autosomal recessive).
ERT (Enzyme Replacement Treatment) is one of the treatments offered to patients with
LPLD.
 (a) What procedure is followed by ERT?
(b) What could be one possible drawback of ERT?
(c) How can LPLD be treated using biotechnology? Elaborate.
3. On spraying Bacillus thuringiensis on an infected cotton crop field the pests are killed by
the toxin, however the toxin although produced by the bacteria does not affect it. Give
reason.
4. Explain the role of restriction endonucleases, gel electrophoresis, selectable marker in
pBR322 in biotechnology.
5. Given below is an autoradiograph of DNA segments, obtained from electrophoresis.

(a) In this autoradiograph, the ladder sequence measures approx 1200 bp, which band
will correspond to 600 bp?
(b) On what basis the fragments get separated like this during gel electrophoresis? Give
one significance of this technique while constructing a recombinant molecule.
6. ‘In some children, ADA deficiency can be cured by bone marrow transplantation or by
enzyme replacement therapy, but both of these approaches are not completely curati
ve. Justify the statement.
7. (a) What is PCR and a primer? (b) What is the denaturation process? (c) Name the bacte
ria from where thermostable DNA polymerase isolated?
8. cry I Ac and cry II Ab are introduced in a plant to prevent infestation by cotton bollworm.
(a) What would be the result of this process? (b) Summarize the action of this gene in
the host it is introduced into.
9. (a) List two advantages of using Bt cotton in comparison to chemical sprays. (b) Does Bt
cotton affect beneficial insects? Discuss.
10. (a) Name the selectable markers in the cloning vector pBR322. Mention the role they
play. (b) Why is the coding sequence of an enzyme β-galactosidase a preferred select
able marker in comparison to the ones named above?
11. Why must a cell be made ‘competent’ in biotechnology experiments? How does the cell
can be made competent? Also state the role of ‘biolistic gun’ in biotechnology experime
nts.
12. A gel electrophoresis was run to show the fragments produced by restriction digestion
with different restriction enzymes. The MWR lane indicates the molecular weight rules.
(a) Construction a restriction map of the double digest of a plasmid.
(b) Explain which fragment will move faster in gel electrophoresis and why?
 13. List any two applications of recombinant DNA technology, each in medical diagnosis and
agriculture.
14. For treating ADA deficiency, gene therapy is carried out. How is this carried out and help
the patient to overcome the disorder? If the gene therapy is performed at embryonic
stages, how will this have a positive effect on the children?
15. State the functions of ori, rop and Hind III sites in the cloning vector pBR322. How does
exonuclease activity differ from endonuclease?
16. A farmer grows two varieties of crop in fields ‘A’ and ‘B’. He grew normal tobacco plant
in field A and GM crops in field B. He observed that nematode attacked only in field A
whereas crops in field B remains unaffected.
(a) Name the nematode responsible for affecting the crops of field ‘A’. (b) How did the
crops of field ‘B’ remain unaffected? (c) Name the vector used to introduce specific
genes into the host plant.

4 marks case based questions/ 5 marks questions

1. The structure below shows pUC18 which is similar to pBR322 in its function. How
ever, they differ in some of their restriction sites and number of ori. The ori number
for pBR322 is approximately 20.

(a) How are pUC18 and pBR322 used in biotechnological studies?

Or
What will be the impact if ori in the above structure gets damaged?
(b) The lac z gene has many recognition sites. Study the segment of DNA given
below and answer the questions
5’….. ATC GTA AAG CTT CAT…..3’
3’….. TAG CAT TTC GAA GTA…..5’
Applying your knowledge of palindrome sequences identify and mark the
possible region where the restriction enzyme X will act.
 Restriction enzyme Y was used to extract gene of interest from a plant. This
gene needs to be inserted in the given DNA segment which has been treated
with restriction enzyme X. Will there be a successful recombination? Explain
with a reason.
(c) Which one of the two (pUC18 and pBR322) would you prefer for biotechnolo
gical studies? Justify.
2. (a) List the operational guidelines that must be adhered to so as to achieve optimi
sation of the bioreactor system. Enlist any four.
(b) Mention the phase of the growth we refer to in the statement “Optimisation of
growth and metabolic activity of the cells”.
(c) Is the biological product formed in the bioreactor suitable for the intended use
immediately? Give reason in support of your answer.
(d) Give the name of the type of bioreactor shown.

List five growth conditions that a bioreactor should provide for obtaining the desired
product.
3. (a) ‘EcoRI’ has played very significant role in rDNA technology. (i) Explain the conven
tion for naming EcoRI. (ii) Write the recognition site and the cleavage sites of this res
triction endonuclease. (b) What are the protruding and hanging stretches of DNA
produced by these restriction enzymes called? Describe their role in formation of
rDNA.
4. (a) GM crops are designed to develop natural resistance from insects & pests. Which
crops are modified using Bacillus thuringiensis? (b) List five advantages of GMOs to a
farmer. (c) Foods derived from genetically modified crops should be tested for possi
ble reactions in people. Explain.
5. (a) Evaluate the efficacy of Golden rice to meet the daily intake of vitamin-A on peo
ple. (b) The GM crops are the best solution to overcome issues developed due to co
nventional breeding. Discuss. (c) List the changes that are done to genetically mod
ified plants.
6. (a) Are you aware of the traits for which genetic modification is carried out in GM
crop plants? (b) The first stage in making a GM plant requires transfer of DNA into a
plant cell. How is this achieved? (c) What is the role of government agencies in the
regulation of all activities related to GMOs and their products?
7. Observe the diagram shown below for the process of humulin production and
answer the questions that follows.
(a) What does label ‘C’ represent and what is its role? Also, identify the label which
represents an enzyme that is utilized for cutting the molecule of DNA.
 (b) What purpose is humulin term used for? Through which bond chains A and B of
humulin are bonded? Which DNA sequence will be introduced in label B to prod
uce humulin?
(c) Name the source from which it was extracted earlier. Why is this no more in use
by diabetic people?
(d) Explain the process of its synthesis by Eli Lilly company.

8. Observe each stage and explain briefly about the process involved in each step.
Stages of rDNA technology Process of rDNA technology
Isolation of genetic material
Cutting of DNA at specific locations
Separation & isolation of DNA fragments
Amplification of gene
Ligation of DNA fragment into a vector
Insertion of rDNA into host
Desired product

9. (a) Which conditions apart from the temperature, pH and substrate are provided by
the bioreactor? (b) What happens in a continuous culture system? (c) Name the
components of a bioreactor. (d) Write the name of two bioreactors. (e) What is the
purpose of sparged stirred tank bioreactor?