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Genetic Engineering
to enable students to-
- get an insight of tools of genetic engineering.
- understand steps involved in the formation of genomic library and fundamentals of gene sequencing.
- know the strategies of genome mapping and analysis of genetic variations.
- understand the techniques involved in gene expression studies.
- know and understand in detail strategies of gene delivery.
Scope and milestones in genetic engineering, Basic tools and techniques used in recombinant DNA technology: Restriction endonuclease, DNA modifying enzymes, cloning vectors: plasmids, bacteriophage, cosmid, phagemids, in vitro construction of vectors, expression vectors. Principle and uses of nucleic acid hybridization. Principle and applications of polymerase chain reaction, in vitro DNA synthesis, in vitro transcription and translation.
DNA and genomic library: m- RNA enrichment, reverse transcription, Linkers, Adaptors, Screening of c DNA and genomic library, Sequencing and mapping: Sequencing vector, fluorescent tagging, Automated DNA sequencing, Pyrosequencing, Restriction mapping and map construction, Application of sequence information for identification of defective genes, Q-PCR.
Genetic and physical mapping, Genome sequencing: genome size, organelle genome, YAC, BAC libraries, strategies of genome sequencing, Analysis of genetic variations: RAPD, RFLP, AFLP and other molecular marker techniques, application of RFLP in forensic studies, disease prognosis, genetic counselling, pedigree analysis etc.
Agrobacterium mediated transformation, electoporation, particle bombardment, microinjection, Gene therapy: Target gene replacement, gene knockout technique, computer aided drug designing, Patenting of cloned life forms. Site directed mutagenesis and protein engineering
DNA and protein microarray technology, RNase protection assay, Reporter gene assay, northern blotting and S1 nuclease assay, Heterologous gene expression in bacteria, yeasts, insects, mammals and plants. codon optimization
1. Molecular Cloning: a Laboratory Manual, J. Sambrook. E.F. Fritsch and T. Maniatis, Cold Spring Harbor Laboratory Press, New York, 2000. 2. DNA Cloning: a practical Approach, D. M. Glover and B.D. Hames, IRL Press Oxford, 1955. 3. Molecular and Cellular Methods in Biology and Medicine, P. B. Kaufman, W. Wu., D. Kim and L.J: Cseke, CRC Press.Florida,1995. 4. An Introduction to Genetic Engineering. Edited by Desmond S.T. Nicholl, Cambridge University Press, February 2002. 5. Manipulation and Expression of Recombinant DNA. Sue Carson and Dominique Robertson, Second edition, Academic Press, December 2005. 6. Principles of Gene Manipulation and Genomics. Primrose and Twyman, (7th edition). Blackwell Publishers, 2006. 7. RNA interference Technology- From basic science to drug development. Edited by Krishnarao Appasani. Forewords by Andrew Fire and Marshall Nirenberg, Cambridge Press, 2005. 8. Methods in Enzymology Vol.152,Guide to Molecular Cloning Techniques, S. L. Berger and A. R. Kimmel, Academic press. Inc. San Dlogo, 1998. 9. Methods in Enzymology Vol.185,Gene Expression Technology, D. V. Gooddol, Academic Press, Inc., San Diego,1990. 10. DNA Science. A First Course in Recombinant Technology, D.A. Mickloss and G.A. Greyer, Cold Spring Harbor Laboratory Press, New York,1990 11. Molecular Biotechnology (2nd Edition), S.B. Primorse, Blackwell Scientific Publishers, Oxford, 1994.
1. Milestones in Biotechnology. Classic papers on genetic Engineering, J.A. Davies and W.S.Roznikolf, Butterworth-Helnemann, Boston, 1992. 2. Route Maps in Gene Technology, M. R. Walker and R.Repley, Blackwell Science Ltd. Oxford, 1997. 3. Genetic Engineering. An Introduction to gene analysis and exploitation in eukaryotes. S. M. Kingsman and A.J. Kingsman, Blackwell Scientific Publications. Oxford, 1998.
