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CHAPTER 11

BIOTECHNOLOGY :PROCESS AND PRINCIPLES

1 MARKS QUESTIONS
1. What is the role of ethidium bromide during during agarose-gel electrophoresis of DNA
fragments?
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2.Mention the role of molecular scissors in rDNA technology.
.
3.Why is it essential to have a selectable marker in cloning vector?
4.Why is the enzyme cellulase used for isolating genetic material from plant cells but not animal
cell?
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5.How is the action of exonuclease different from that of endonuclease ?
.
6.Name the host cell that produces a foreign gene product ? What is the product called ?

How can bacterial DNA be released from the bacterial cell for biotechnology experiments?
8. Mention the uses of cloning vector in biotechnology.
9.Why do DNA fragments move towards anode during gel electrophoresis?
10.In the year 1963 ,two enzymes responsible for restricting the growth of bacteriophage in E.coli
were isolated .How did the enzyme restrict the growth of bacteriophage?
11. Suggest a technique to a researcher who needs to separate fragments of DNA.
12. Mention the role of Restriction Enzymes in Recombinant DNA technology.
13. Write the two components of the first artificial recombinant DNA molecule constructed by
Cohen and Boyer.
2 MARKS QUESTIONS
1.Name the source of the DNA –polymerase used in PCR technique. Mention why it is used.
2. Write any four ways used to introduce a desired DNA segment into a bacterial cell in
recombinant DNA technology experiments.

3. Why is ORIGIN OF REPLICATION (ORI) required to facilitate cloning into a vector?

4. Name a natural source of agarose .Mention one role of agarose in biotechnology.
5. Name two commonly used bioreactors.State the importance of bioreactors.
6. Explain how to find whether an E.coli bacterium has transformed or not ,when a recombinant
DNA bearing ampicillin –resistant genes transformed into it .
7. a) A recombinant vector with a gene of interest inserted within the gene of β-galactosidase
enzyme is introduced into a bacterium .Explain the method that would help in selection of
recombinant colonies from non-recombinant ones.
b) Why this method of selection referred to as insertional inactivation ?
8. Explain the action of the restriction endonuclease EcoRI.
9.Explain the role of Ti plasmid in biotechnology ..

3 MARKS QUESTIONS
1.a)Explain the significance of ‘palindromic nucleotide sequence’ in the formation of recombinant
DNA.
(b) Write the use of restriction endonuclease in the above process.
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2.Describe the roles of heat, primers and the bacterium Thermus aquaticus in the process of
PCR.
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3.a) Name the selectable marker in the cloning vector pBR322? Mention the role they play.
b)Why is the coding sequence of an enzyme β-galactosidase a preferred selectable marker in
comparison to the named above?
4.a)Why must cell be made competent in biotechnology experiments? How does the calcium ion
help in doing so?
b) State the role of biolistic gun in biotechnology experiment.
5.Suggest and describe a technique to obtain multiple copies of a gene of interest in vitro.

6.Why does the ‘insertional inactivation’ method to detect recombinant DNA is preferred to
‘antibiotic resistance’ procedure ?
7.Explain the role of the enzyme EcoRI in recombinant DNA technology.
8.Mention the role of (i) selectable marker, (ii) Ori and (iii) rop in E. coli cloning vector pBR322.
.
9.Draw a diagram of a typical agarose gel electrophoresis showing migration of undigested and
digested sets of DNA fragments. Label (a) the digested and undigested DNA fragments, (b) Anode
and cathode ends of the plate. Mention the role of electrophoresis in biotechnology.

Isolate and analyse the DNA molecules which are cut by restriction enzymes which are used for
cloning purpose.
11.How does β-galactosidase coding sequence act as a selectable marker ? Explain. Why is it a
preferred selectable marker to antibiotic resistance genes ?
12.How can a bioreactor be made to function at optimal state in order to obtain a desired foreign
gene product ? Explain.
13.Given below is the diagram of agarose gel kept under UV light:

(a) Mention the positive and negative terminals.

(b) What is the charge carried by DNA molecule and how does it help in its
separation?
(c) How are the separated DNA fragments finally isolated?
Ans. after adding ethidium bromide, the DNA fragments are viewed under UV light and viewed as
orange coloured bands. Then the desired fragments are separated by a knife.
13.(a) In pBR322, foreign DNA has to be introduced in tetR region. From the restriction enzymes
given below, which one should be used and why: PvuI, EcoRI, BamHI
(b) Give reasons, why the other two enzymes cannot be used.
.
5 MARKS QUESTIONS
1.a)Why are the engineered vectors preferred by biotechnologists for transferring the desired
genes into another organism?
b)Explain how do ‘ORI’ “selectable markers” and cloning sites facilitate cloning into vector.
2.i)Describe the characteristics that a cloning vector must possess.
ii)Why DNA cannot pass through the cell membrane ? Explain. How a bacterial cell is made
‘competent’ to take up recombinant DNA from the medium?
ii)DNA is hydrophilic molecule and cell membrane is made up of phospholipid

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18-12-2017 SABITABRATA MANDAL