M.Sc-M.Sc BioTechnology 2nd Sem BT - 21 : Genetic Engineering(University of Pune, Pune-2013)

                            M.Sc. (Semester - II) BIOTECHNOLOGY

SEAT No. :

[Total No. of Pages : 2

BT - 21 : Genetic Engineering

(2008 Pattern)

Time : 3 Hours]                                                                                                     [Max. Marks : 80

Instructions to the candidates:

Attempt not more than 5 questions of which at least 2 questions must be from each

section.

Answers to the two sections should be written in separate books. Neat diagrams must be drawn wherever necessary. Figures to the right indicate full marks.

SECTION - I

Q1) Write short notes on :

a) Regulation of copy number in plasmids. b) Chimeric construct.

c) Synthetic promoters used in expression vectors. d) Construction of cDNA library.

Q2) a) Comment on the difference in strategies used for cloning prokaryote and

eukaryote genes.                                                                                                  [8]

b) Explain the mechanism and applications of lac operon in expression of

industrially important products.                                                               [8]

Q3) a) Describe the methods used for screening and selection of transformed

cells in gene library construction.                                                             [8]

b) i) Give the importance of topoisomerase & ligase enzyme.

ii) With suitable example describe a typical yeast expression vector.

[8]

Q4) a) Give a comparative account between plasmid and phage as DNA carrier with

respect to host, insert size, entry in host, efficiency and application.            [8]

P.T.O.

b) i)         Write a brief note on restriction enzymes, its types and role in genetic

modification.

ii) Construct a restriction map for a 8.9 kb circular plasmid which is

singly and doubly digested with 3 restriction enzymes, EcoRI, Bam HI and Hind III.

Following bands were obtained :

Eco RI – 8.9 kb

Bam HI – 6 kb & 2.9 kb

Hind III – 8.9 kb

EcoRI + Bam HI – 6.0 kb, 2.4 kb & 0.5 kb

EcoRI + Hind III – 7.4 kb & 1.5 kb

Bam HI + Hind III – 5 kb, 2.9 kb & 1 kb

EcoRI + Bam HI + Hind III – 5 kb, 2.4 kb, 1 kb, & 0.5 kb            [8]

SECTION - II

Q5) a) What are Co - dominant markers? Explain the development of

miciosatellite markers.                                                                                        [8]

b) Explain the technology used for creating the cloned sheep ‘Dolly’. Give,

reason for early death of Dolly at the age of 6 (normal sheeps live upto to

12 years).                                                                                                              [8]

Q6) Write an assay on any two methods employed for transgenic plant

development. Add a note on biosafety precautions taken while releasing

transgenic plants.                                                                                                 [16]

Q7) a) Explain the recipe for a typical polymerase chain reaction, commenting

on the importance of each ingredient.                                                     [8]

b) Give the principle of Sanger’s di - deoxy method for sequencing a gene.

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