Code: 9A23504

B.Tech III Year I Semester (R09) Regular & Supplementary Examinations December/January 2013/14

GENETIC ENGINEERING
(Biotechnology)
Time: 3 hours Answer any FIVE questions All questions carry equal marks ***** 1. How many sigma factors are present in prokaryotic gene expression system? And explain how do sigma factors bind to promoters. Write note on the following with suitable examples: (a) Promoter. (b) Enhancer elements. Write an essay on transposable elements in bacteria. What problems may be encountered when using Lambda as a cloning vector? Briefly describe how you would prepare a genomic library in a lambda replacement vector. Describe about Microarray technology and how it is used in gene expression analysis. Describe the Sanger method for determining the nucleotide sequence of DNA and the two different methods that can be used to detect DNA fragments generated in the sequencing reaction. Discuss the polymerase chain reaction, including the method itself and the components of the PCR reaction mixture. Finally describe two examples of the uses to which PCR can be put in genetic engineering. Explain the advantages and limitations of gene therapy. Max. Marks: 70

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Code: 9A23504

B.Tech III Year I Semester (R09) Regular & Supplementary Examinations December/January 2013/14

GENETIC ENGINEERING
(Biotechnology)
Time: 3 hours Answer any FIVE questions All questions carry equal marks ***** 1. 2. Write an essay on mechanism of gene regulation in prokaryotes. Write an essay on the structure of eukaryotic genes. You should consider both the regulatory and coding regions. What is a plasmid? Discuss the properties of modern plasmid vectors that make them suitable for the selection of recombinants and for the cloning and expression of foreign DNA. Describe the general properties of restriction enzymes and their use in Genetic engineering. Describe the processes involved in creating a gene library from eukaryotic nucleic acids. You should make clear the difference between a cDNA library and a genomic library. Explain the parameters involved in designing of primers. Compare the relative advantages and disadvantages of the following molecular markers: RAPD, RELP, SSR Comment on the following: (a) Transient and stable expression. (b) Phospholipids as gene-delivery vehicles. (c) Electroporation. ***** Max. Marks: 70

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Code: 9A23504

B.Tech III Year I Semester (R09) Regular & Supplementary Examinations December/January 2013/14

GENETIC ENGINEERING
(Biotechnology)
Time: 3 hours Answer any FIVE questions All questions carry equal marks ***** 1. 2. 3. 4. What is operon? Explain the structure and mechanism of Lac operon in detail. Write an essay on repetitive DNA with suitable illustrations. What is retrotransposons? Explain the mechanism and types in detail. Define each of the following, describing how they are used in the cloning and analysis of DNA: (a) Restriction endonucleases. (b) DNA ligase. Describe: (a) The principles that underlie southern blotting. (b) The practical steps necessary to obtain a southern blot and (c) Provide and explanation for the following result: Digestion of a gene with EcoRI gives four fragments with sizes of 3.01 kb, 2.7kb, 2.0kb and 0.8 kb. Southern blotting of EcoRI digested genomic DNA with full length cDNA derived from this gene detects fragments with sizes of 3.1 kb, 2.7 kb and 0.8 kb. How would you explain this? Comment on the following: (a) Key factors affecting the PCR. (b) Applications of PCR. What is molecular marker? Explain any one hybridization based molecular marker in detail. Describe about the Baculovirus expression system with suitable diagram. Max. Marks: 70

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Code: 9A23504

B.Tech III Year I Semester (R09) Regular & Supplementary Examinations December/January 2013/14

GENETIC ENGINEERING
(Biotechnology)
Time: 3 hours Answer any FIVE questions All questions carry equal marks ***** 1. 2. 3. Describe the repressors involved in gene regulation and expression in prokaryotes. Write an essay on the structure and function of eukaryotic gene regulatory proteins. Write about the following with suitable diagram. (a) Insertional inactivation. (b) Homologous recombination. Draw the vector map of the following: (a) pBR322. (b) pUC 19. (c) Any expression vector with fusion protein. What are the differences between a genomic library and a cDNA library? In your answer, you should refer to the starting material, the types of vectors that can be used to make the libraries and how the libraries are made. Write detail note on RT PCR and multiplex PCR with suitable diagram. Explain the AFLP technique and their relative merits in detecting DNA polymorphisms. How you express high amount of foreign protein in E.coil, explain with suitable example? ***** Max. Marks: 70

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