Biotechnology

Exercise-II

SECTION A: KVPY PYQs

1. The restriction endonuclease EcoRI recognizes and cleaves DNA sequence as shown
below-
5’ – GAATTC – 3’
3’ – CTTAAG – 5’
What probable number of cleavage sites can occur in a 10 kb long random DNA
sequence?
[KVPY 2012 SB/SX]
(A) 10
(B) 2
(C) 100
(D) 50

2. Eco RI and Rsa I restriction endonucleases require 6 bp and 4 bp sequences respectively

for cleavage. In a 10 kb DNA fragment, how many probable cleavage sites are present for
these enzymes?
[KVPY 2013 SB/SX]
(A) 0 Eco RI and 10 Rsa I
(B) 1 Eco RI and 29 Rsa I
(C) 4 Eco RI and 69 Rsa I
(D) 2 Eco RI and 39 Rsa I

3. A scientist has cloned an 8 kb fragment of a mouse gene into the Eco RI site of a vector
of 6 kb size. The cloned DNA has no other Eco RI site within. Digestions of the cloned
DNA are shown below.
 Which one of the following sets of DNA fragments generated by digestion with both Eco
RI and Bam HI as shown in (iii) is from the gene?
[KVPY 2013 SB/SX]
(A) 1 kb and 4 kb
(B) 1 kb and 2.5 kb
(C) 1 kb and 3 kb
(D) 1 kb and 3.5 kb

4. A circular plasmid of 10,000 base pairs (bp) is digested with two restriction enzymes, A
and B, to produce a 3000 bp and a 2000 bp bands when visualized on an agarose gel.
When digested with one enzyme at a time, only one band is visible at 5000 bp. If the first
site for enzyme A (A1) is present at the 100th base, the order in which the remaining sites
(A2, B1, and B2) are present is:
[KVPY 2014 SB/SX]
(A) 3100, 5100, 8100
(B) 8100, 3100, 5100
(C) 5100, 3100, 8100
(D) 8100, 5100, 3100

5. Restriction endonucleases are enzymes that are used by biotechnologists to

[KVPY 2015 SA]
(A) Cut DNA at specific DNA sequences
(B) Join fragments of DNA
(C) Digest DNA from the 3’ end
(D) Digest DNA from the 5’ end

6. Genomic DNA is digested with Alu I, a restriction enzyme which is a four base-pair
cutter. What is the frequency with which it will cut the DNA assuming a random
distribution of bases in the genome?
[KVPY 2015 SA]
(A) ¼
(B) 1/24
(C) 1/256
(D) 1/1296
7. In biotechnology application, a selectable marker is incorporated in a plasmid
[KVPY 2015 SB/SX]
(A) To increase its copy number
(B) To increase the transformation efficiency
(C) To eliminate the non-transformants
(D) To increase the expression of the gene of interest

8. Consider the linear double-stranded DNA shown below. The restriction enzyme sites and
the lengths are demarcated as shown. This DNA is completely digested with both Eco RI
and Bam HI restriction enzymes. If the product is analyzed by gel electrophoresis, how
many distinct bands would be observed?
[KVPY 2015 SB/SX]

(A) 5
(B) 2
(C) 3
(D) 4

9. How many bands are seen when immunoglobulin G molecules are analyzed on a sodium
dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing
conditions?
[KVPY 2015 SB/SX]
(A) 6
(B) 1
(C) 2
(D) 4

10. In the presence of glucose and lactose, Escherichia coli utilizes glucose. However,
lactose also enters the cells because
[KVPY 2016 SA]
(A) Low level of permease is always present in the cell
(B) It uses the same transporter as glucose
(C) It diffuses through the bacterial cell membrane
(D) It is transported through porins
11. Which ONE of the following is NOT essential for Polymerase Chain Reaction (PCR)?
[KVPY 2017 SB/SX]
(A) Restriction enzyme
(B) Denaturation of DNA
(C) Primers
(D) DNA polymerase

12. Which ONE of the following combinations of molecular masses of polypeptides are
obtained from purified human IgM when analyzed on sodium dodecyl sulfate
polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions?
[KVPY 2017 SB/SX]
(A) 55 kDa, 15 kDa
(B) 70 kDa, 25 kDa, 15 kDa
(C) 55 kDa, 25 kDa
(D) 155 kDa

13. The schematic below describes the status of lac operon in the absence of lactose. Which
ONE of the following happens when lactose is present in the cell?

[KVPY 2017 SB/SX]

i
(A) Lactose binds to P and stops the transcription of i.
(B) Lactose is converted to allolactose, which binds to Plac and results in the displacement
of the repressor from O.
(C) Lactose is converted to allolactose, which binds to the repressor protein and prevents
its interaction with O.
(D) Lactose has no effect on the status of the lac operon.
14. Bacterial plasmids are genetic entities that
[KVPY 2017 SB/SX]
(A) Are non-transferable to the same bacterial species.
(B) Are capable of independent replication.
(C) Have RNA as genetic material.
(D) Always require integration in the genome for their replication.

15. As indicated in the gel image, lanes X and Y represent samples obtained from a circular
plasmid DNA after complete digestion using restriction enzyme X or Y with different
recognition sites, respectively. How many sites for X and Y are present in the plasmid
(sizes of the bands in kilobase pairs (kb) is shown)?

[KVPY 2017 SB/SX]

(A) 1 for X, 1 for Y
(B) 2 for X, 1 for Y
(C) 1 for X, 2 for Y
(D) 2 for X, 2 for Y

16. Bt toxin produced by Bacillus thuringiensis does not kill the producer because the toxin
is
[KVPY 2018 SB/SX]
(A) In an inactive protoxin form
(B) Rapidly secreted outside
(C) Inactivated by an antitoxin
(D) In unfolded form

17. Which ONE of the following primer pairs would amplify the fragment of DNA given
below?
5’ – CTAGTCGTCGAT – (N)300 – GACTGAGCTGAGCTG – 3’
3’ – GATCAGCAGCTA – (N)300 – CTGACTCGACTCGAC – 5’
[KVPY 2018 SB/SX]
 (A) 5’ – CTAGTCGTCGAT – 3’ and 5’ – GACTGAGCTGAGCTG – 3’
(B) 5’ – CTGACTCGACTCGAC – 3’ and 5’ – CTAGTCGTCGAT – 3’
(C) 5’ – CTAGTCGTCGAT – 3’ and 5’ – CAGCTCAGCTCAGTC – 3’
(D) 5’ – CTAGTCGTCGAT – 3’ and 5’ – GTCGAGTCGAGTCAG – 3’

18. Which of the following elements is NOT directly involved in transcription?

[KVPY 2019 SB/SX]
(A) Promoter
(B) Terminator
(C) Enhancer
(D) OriC

19. Which ONE of the following statements is CORRECT about the mechanism of action of
penicillin?
[KVPY 2020 SB/SX]
(A) It inhibits transcription
(B) It hydrolyzes cell wall
(C) It inhibits cell wall biosynthesis
(D) It inhibits translation

SECTION B: NEST PYQs

20. A pure culture of Escherichia coli was streaked on a plate to get single isolated colonies.
The plasmid DNA from two such colonies was individually isolated. The DNA was then
individually treated with Eco RI (P and Q) or double digested with Eco RI + Hind III (P’
and Q’). The digested DNA was electrophoresed on an agarose gel and the pattern of the
linear DNA so obtained is depicted in the figure below. Patterns P and P’ are obtained for
DNA from colony 1 and are as expected for the original DNA sequence of the plasmid.
The best reason for the differing pattern in Q’ is because the plasmid DNA from colony 2
has a mutation _________.
[NEST 2020]
 (A) Resulting in another Hind III site
(B) At its original Eco RI site
(C) At its original Hind III site
(D) Resulting in another Eco RI site

21. A drug P inhibits DNA replication in the bacterium E. coli by specifically blocking the
catalytic site of DNA polymerase activity. That the same drug P can be used to treat
cancers in humans can be explained from the fact that ______________.
[NEST 2020]
(A) Mechanism of DNA polymerase is majorly conserved
(B) Both DNA polymerases require RNA as a primer
(C) Both cancer and bacterial cells divide rapidly into large numbers
(D) DNA replication is bi-directional in both the organisms

22. A polymerase chain reaction (PCR) for the amplification of DNA fragments involves
multiple cycles of thermal denaturation, annealing, and extension steps in each cycle.
Because of the heat denaturation step, a thermostable polymerase is required for
performing PCR. An experimenter had all the resources except the thermostable DNA
polymerase to perform a PCR. The experimenter can use ____________ to substitute for
thermostable DNA polymerase.
[NEST 2020]
(A) Escherichia coli DNA polymerase added before denaturation step of every PCR cycle
(B) Escherichia coli DNA polymerase, added after extension step of every PCR cycle
(C) Excess amount of Escherichia coli DNA polymerase added once at the beginning of
the reaction
(D) Escherichia coli DNA polymerase, added after annealing step of every PCR cycle
23. Barbara McClintock observed non-Mendelian pigmentation patterns in maize (figure 1)
mediated by transposons, as also observed in Petunia (figure 2 and 3). These somatic
changes mere inheritable as evident from the clonal -patterns of pigmentation observed
both in seeds and flowers.

As leaves grow, ‘jumping genes’ get activated and insert themselves into
chlorophyll-producing genes, thereby disrupting and inactivating them. This is typically
seen as leaf pigment variegation (P and Q). Sometimes, the transposon jumps out from
where it was inserted, restoring the chlorophyll gene function. Observing the leaf
variegation patterns in P and Q, choose the correct statement(s).

[NEST 2020]
(A) Only Q shows clonal propagation of the variegated phenotype.
(B) Only P largely indicates that the transposon can jump out of the chlorophyll gene,
restoring its function.
(C) Tissue culture propagated plants from green tissue of leaf Q will show no variegation.
(D) Both P and Q show sporadic events of transposon-mediated variegation.

24. Escherichia coli strains (I-IV) with mutations in the lac operon were studied for
β-galactosidase activity, followed by measuring its mRNA and protein levels (Graphs).
The arrow indicates the time of addition of a compound (namely IPTG) that induces the
lac operon. Based on the results, the correct option(s) is(are)
 [NEST 2021]
(A) In strain-I, the lac operon is not regulated and β-galactosidase is inactive.
(B) In strain-II, the lac operon is not regulated and β-galactosidase is inactive.
(C) In strain-IV, the lac operon is not regulated and β-galactosidase protein is not
produced.
(D) In strain-III, the lac operon is regulated and β-galactosidase is active.

25. A student wants to construct a plasmid ‘BacLight’, that when transformed into a
bacterium will produce light IF AND ONLY IF the sugar lactose is present in the
medium. For this, she uses the following DNA sequences: promoter of the repressor (Pi),
promoter of lacZ (Pz), operator (O) and repressor gene lacI from the lactose operon; and
a structural gene X, which encodes a protein that produces light. The different DNA
elements (II - V) and a vector (I) were digested with restriction enzymes and ligated to
produce the plasmid. The DNAs used along with restriction enzyme sites at their ends are
given below. Slash (/) in the sequence represents the point at which the restriction
enzyme cuts the DNA.
I : 5′ —(G/GATCC) –Vector –(T/CTAGA) —3′
II : 5′ —(T/CTAGA) –Pi –lacI –(G/GATCC) —3′
III : 5′ —(T/CTAGA) –Pz –(G/CTAGC) —3′
IV : 5′ —(G/CTAGC) –O –(A/CTAGT) —3′
V : 5′ —(A/CTAGT) –Gene X –(T/CTAGA) —3′
Choose the correct sequence of DNAs that can possibly be ligated to produce the plasmid
‘BacLight’.
[NEST 2017]
(A) Vector –Pi –lacI –Pz –O –Gene X –Vector
(B) Vector –Pz –O –Gene X –Pi- lacI –Vector
(C) Vector –Pz –Gene X –O –Pi- lacI –Vector
(D) Vector –Pz –O –Pi –lacI –Gene X –Vector
26. E. coli is a bacterium routinely used in cloning experiments and can be grown on
artificially synthesized media in the laboratory. One of the strains of this bacterium, XL-1
Blue, is naturally resistant to the antibiotic tetracycline. A genetic transformation
experiment was performed to introduce a plasmid that encodes an ampicillin resistance
gene into this strain. Following transformation, an equal number of cells were grown
independently on two types of growth media - (A) growth medium with tetracycline
alone and (B) growth medium with tetracycline and ampicillin. The number of colonies
that grew on medium A was much higher than that obtained on medium B. Which of the
following statement(s) explain the above observation?
[NEST 2012]
(A) The transformation efficiency of cells is lower than their viability.
(B) Untransformed cells also grew on growth medium “A”.
(C) Tetracycline was inhibitory to the growth of bacterial cells.
(D) Ampicillin did not allow transformed cells to grow on growth medium “B”.

27. Which of the following pairs is an INCORRECT match?

[NEST 2016]
(A) Transcriptional repression — gene regulation
(B) Post-translational modifications — phosphorylation
(C) Cytoplasm — exon splicing of eukaryotic pre-mRNA
(D) Enhancers — increased transcription

28. An experimenter ligated cDNA of p53 gene into the multiple cloning site (MCS) of a
vector whose map is shown below. The ligated vector was transformed into E. coli cells, and
transformants were initially selected on an agar plate containing antibiotic P. The bacterial
colonies obtained on this plate were further propagated sequentially on antibiotic Q and then
antibiotic R by replica plating.
The correct sequence(s) of P, Q and R to be used to select E. coli colonies with p53 cDNA
cloned into the MCS is/are:
[NEST 2022]
(A) Ampicillin → Chloramphenicol → Tetracycline
(B) Chloramphenicol → Tetracycline → Ampicillin
(C) Ampicillin → Tetracycline → Chloramphenicol
(D) Tetracycline → Chloramphenicol → Ampicillin

29. Primers P, Q, R and S have binding sites on the template DNA as depicted in the figure
below.

In a polymerase chain reaction (PCR) using all four primers together, the possible number of
amplified products of different sizes is:
[NEST 2022]
(A) 2
(B) 4
(C) 3
(D) 1

30. Schematic P shows restriction enzyme site mapping on a DNA fragment containing gene w
that codes for protein W. Schematic Q shows a plasmid-based vector map and restriction enzyme
recognition site sequences in the multiple cloning site (MCS). The oblique lines in the sequences
next to each restriction enzyme name show the cleavage sites of the respective restriction
enzymes. If the goal of an experimenter is to clone the gene w into the vector to express the
protein W, the correct restriction enzyme pair(s) from the options below that can be used to
digest the DNA fragment as well as the vector is/are:
 [NEST 2022]
(A) HindIII and Apal
(B) HindIII and EcoRV
(C) EcoRV and ApaI
(D) BamHI and EcoRV

SECTION C: IAT PYQs

31. As opposed to DNA replication within the cell, discontinuous synthesis of DNA does NOT
occur in a polymerase chain reaction (PCR). Why?
[IAT 2022]
(A) Denaturation step in PCR substitutes for the replication fork and Taq polymerase extends
DNA only in the 5′ to 3′ direction
(B) The replication fork is formed and Taq polymerase extends DNA in both 3′ to 5′ and 5′ to
3 ′ direction
(C) Denaturation step in PCR substitutes for the replication fork and Taq polymerase extends
DNA only in the 3′ to 5′ direction
(D) The replication fork is formed and DNA ligase activity of the Taq polymerase joins the
discontinuous fragments.
32. Which one of the following is an example of palindromic DNA sequence?
[IAT 2021]
(A) GAATTC
CTTAAG
(B) GACTTC
CTGAAG
(C) GAAGTC
CTTCAG
(D) GACCAG
CTGGTC

33. The map of a 4000 base pair (bp) plasmid DNA marking the locations of different restriction
enzyme cut sites is shown in the figure below. The numbers in brackets indicate the base pair
positions where the enzymes cut. This plasmid is completely digested first with the combination
of restriction enzymes PstI and HindIII, and then with EcoRI and SalI. The final digested
plasmid is analyzed by agarose gel electrophoresis.

[IAT 2021]
Which one of the following options correctly represents the bands sizes (in bp) obtained on the
gel?
(A) 220, 540, 900 and 2340
(B) 220, 540, 1200 and 2040
(C) 540, 760, 1200 and 1500
(D) 320, 540, 800 and 2340
34. DNA fragments of 100 base pairs (bp), 300 bp and 500 bp were separated by agarose gel
electrophoresis. Pick the correct arrangement of the fragments separated on the gel in the
increasing order of their migration from the wells.
[IAT 2018]
(A) 500 bp < 300 bp < 100 bp
(B) 100 bp < 300 bp < 500 bp
(C) 100 bp 300 bp > 500 bp
(D) 500 bp> 300 bp > 100 bp

35. A gene is inserted into the PvuII site of the cloning vector (given below) and transformed
into E. coli cells. Which one of the following statements is then true?

[IAT 2018]
(A) Recombinants can be selected by plating on ampicillin containing medium
(B) Recombinants can be selected by plating on tetracy- cline containing medium.
(C) Recombinants cannot be selected by plating either on ampicillin or tetracycline
containing medium
(D) Recombinants can be selected by plating simultaneously on ampicillin and tetracycline
containing medium.
36. Provided below are recognition sequences for restriction enzymes 1 (RE1), 2 (RE2) and 3
(RE3). Arrows indicate the positions where the enzymes digest on the two strands. Which of the
following can the RE1 digested DNA ligate to?

[IAT 2017]
(A) All three RE1, RE2 and RE3 digested DNA
(B) Only to RE1 digested DNA
(C) Only to RE1 and RE3 digested DNA
(D) Only to RE1 and RE2 digested DNA
 Biotechnology
Exercise-II

SECTION A: KVPY PYQs

1.B
2.D
3.C
4.A
5.A
6.C
7.C
8.C
9.C
10.A
11.A
12.B
13.D
14.B
15.B
16.C
17.C
18.D
19.C

SECTION B: NEST PYQs

20.A
21.A
22.D
23.B,D
24.C,D
25.B
26.A,B
27.C
28.B.D
29.B
30.A,C