GENETIC ENGINEERING

Paper Code: 
BTE 402
Credits: 
03
Contact Hours: 
45
Objective: 

This paper will enable students to-

  • learn the basics of Genetic Engineering
  • study different cloning and expression vectors
  • study the enzymes used in Genetic Engineering
  • learn the procedures involved in isolation of genomic and nuclear DNA
8.00
Unit I: 
Introduction to Genetic Engineering

Scope of Genetic Engineering, Milestones in Genetic Engineering, Types of cloning vectors viz. Plasmids, cosmids, ssDNA Phages, Yeast cloning vectors, Animal viruses, Ti plasmids and Cauliflower Mosaic Virus.

10.00
Unit II: 
Plasmids and its various types

Structural and Funtional Organization of Plasmids, Plasmid Replication, Stringent and Relaxed Plasmids, Incompatibility of Plasmid Maintenance. Lambda Phage as a natural in vivo vector, in vitro construction of Lambda vector.

10.00
Unit III: 
Enzymes in Genetic Engineering

DNA Polymerase, Polynucleotide kinase, T4 DNA ligase, Nick translation system, Terminal deoxynucleotidyl transferase, Reverse transcriptase Restriction endonucleases.

9.00
Unit IV: 
Genome Analysis and Sequencing

Isolation of genomic and nuclear DNA: DNA digestion and restriction fragment analysis and sequencing by chemical, enzymatic and automated DNA sequencing methods.

9.00
Unit V: 
RDT and Hybridization techniques

Selection of rDNA clones and their expression products:
Direct and indirect methods. Drug resistance, gene inactivation, DNA hybridization, colony hybridization and in-situ hybridization (Southern, Northern and Dot blots and immunological techniques Western blotting).

ESSENTIAL READINGS: 
  1. S.B.Primorse, (1994) Molecular Biotechnology (2nd Edn.), Blackwell Scientific Publishers, Oxford.
  2. M.R.Walker and R.Repley, (1997). Route Maps in Gene Technology, Blackwell Science Ltd.Oxford.
  3. T.A. Brown ,(2001). Gene Cloning and DNA Analysis. An Introduction. Blackwell Science.
REFERENCES: 
  1. Sambrook J., Fritsch E.F and Maniatis T. (2000) Molecular Cloning: a Laboratory Manual,  Cold Spring Harbor Laboratory Press, New York.
  2. Glover D.M. and Hames B.D. (1955) DNA Cloning: a practical Approach, , IRL Press Oxford.
  3. Kaufman P.B., Kim W.Wu. D.and Cseke L.J, (1995). Molecular and Cellular Methods in Biology and Medicine, CRC Press. Florida,.
  4. Berger S.L. and Kimmel A.R. (1998). Methods in Enzymology, Guide to Molecular Cloning Techniques, Academic press. San Diego,
  5. Gooddol D.V. (1990). Methods in Enzymology, Gene Expression Technology, Academic Press, San Diego.
  6. Mickloss D.A.and Greyer G.A. (1990). DNA Science. A First Course in Recombinant Technology, ,Cold Spring Harbor Laboratory Press, New York,
  7. Davies J.A and Roznikolf W.S. (1992). Milestones in Biotechnology. Classic papers on genetic Engineering, Butterworth-Helnemann, Boston,.
  8. Kingsman S.M. and Kingsman A.J (1998). Genetic Engineering. An Introduction to gene analysis and exploitation in eukaryotes. Blackwell Scientific Publications, Oxford, 1998.
Academic Year: 
2014-2015
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