Thursday, 21st November, 2002

Time: 2.30 – 5.30 p.m.

              Paper: BT- 301. Recombinant DNA technology and  applications.                                                                   Total marks: 70

 

            Note:             Answer sections I and II in separate answer books

                        Numbers within parenthesis indicate marks

 

 

SECTION – I

 

1.a)      Discuss the steps involved in the isolation of plasmid DNA. How does it differ from isolation of Bacteriophage DNA?                  (7)

   b)      Write briefly on the role of DNA ligases in cloning.            (3)

 

OR

 

1.         Outline the salient characters of Restriction endonucleases type I, II,

and III. Explain why   type I and III enzymes are not utilized in cloning?                                                     (10)

 

2.a)      Discuss the advantages and disadvantages of viral vectors over                

                              plasmid vectors.                                                                                    (5)                             b)     Describe the basic characters of  Adenovirus based vectors. What are

                              the advantages of this vector in cloning?                                                (5)

 

OR

 

                  2. a)    Explain the role of the following in a PCR experiment.

                              i) Template DNA                ii) Primers   iii) DNA polymerase (7)

b)        In a PCR experiment, the forward and reverse primers had 50% GC content. What would be the ideal annealing temperature for these primers? Why?                                                 (3)

 

3.        Write short notes on any two:                       (2x5 = 10)

            i) l ZAP vectors            ii) Random primer labeling

           iii) Southern blotting      iv) Colony hybridization.

 

 

SECTION – II

 

4.a)        Distinguish screening and selection. Explain any one genetic method for the selection of recombinants.                                        (6)

   b)   Describe electroporation and its uses in brief.                        (4)

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

4.a)        Distingu

OR

 

4. a)  Give an account of differential screening. What are its applications in

          molecular biology.                                         (6)

   b) Write a short account on ‘in vitro’ packaging.             (4)

 

 

                5.      Write briefly on any two of the following:                                  (2x5 = 10)

i)                    Subtractive DNA cloning                       

ii)                   Visual selection for recombinants

iii)                 Applications of PCR in medicine

iv)                 Particle bombardment

 

 

6.a)  Outline the essential and desirable properties of DNA polymerases used   in sequencing by Sanger’s method.                                                     (7)

                    b) Describe why Maxam – Gilberts method is not amenable for

 automation?                                                                                          (3)

 

OR

 

6.a)   Give a detailed account of the problems caused by the E.coli host in foreign gene expression.                                                                 (7)

   b)   What are search engines? Describe their role in data collection and data mining in Biotechnology.                                                                        (3)

 

7.a)   With the help of examples, discuss the application of biotechnology for the betterment of crops.                                                             (3)

b)     Explain the biochemical mechanism for higher production of Fumaric acid using Rhyzopus oryzae. Discuss the future research on Fumaric acid production.                                                                                                (4)

c)      Explain: gene for gene theory for recognition of pathogenic microorganisms of plants.                                                                        (3)

 

OR

 

7.a)   Describe possible approaches to stabilize transgene expression in plants.

(4)           

b)     Discuss the pros and cons of using microbial cells and mammalian cells

respectively for the production of recombinant proteins.          (4)

c)       Give any two examples of production of biopharmaceuticals by recombinant DNA technology.                                                          (2)

 

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