18BT56 Genetic Engineering &Applications syllabus for BT

VECTORS & ENZYMES IN GENETIC ENGINEERING:

Vectors in rDNA technology, salient features of vectors, types of vectors-plasmids, cosmids, phagemids and viruses. Construction of rDNA & vectors (BAC, Blue script and YAC). Exonucleases and Restriction Endonculeases: classification, mode of action. Enzymes in modification - Polynucleotide phosphorylase, DNase, Methylases, phosphatases, polynucleotide Kinase, Ligases, RNase and their mechansim of action

NUCLEIC ACID HYBRIDIZATION, AMPLIFICATION & CONSTRUCTION OF LIBRARIES:

Methods of nucleic acid detection, polymerase chain reaction (PCR), variants of PCR and applications, methods of nucleic acid hybridization, Southern, Northern & Western hybridization techniques & applications. Isolation of nucleic acids (DNA & RNA). Isolation of plasmids, construction of genomic and cDNA libraries, purification, screening and preservation

METHODS OF GENE/DNA TRANSFER:

Overview & classification of gene transfer techniques in plants, animals and microbes – Transformation, stable & transient transformation, transfection, electroporation, microinjection, liposome mediated gene transfer, transfection of DNA by calcium phosphate coprecipitation, gene gun method. Agrobacterium-mediated gene transfer in plants – Ti & Ri plasmids: structure and functions, Ti plasmid based vectors – advantages, disease control of Agrobacterium tumefaciens. Chloroplast transformation & its applications.

TRANSGENIC SCIENCE IN GENETIC IMPROVEMENT

Transgenic science in plant improvement, biopharming – plants as bioreactors, transgenic crops for increased yield, resistance to biotic and abiotic stresses. Techniques of gene mapping in plants. Marker-assisted selection and breeding for improvement. Transgenic science for animal improvement, biopharming - animals as bioreactors for recombinant proteins, Gene mapping in farm animals. Marker-assisted selection and genetic improvement of livestock.

OTHER APPLICATIONS & GENE THERAPY

Microbial biotechnology - Genetic manipulation, engineering microbes for the production of antibiotics, enzymes, Insulin, growth hormones, monoclonal antibodies, clearing oil spills. Introduction to gene therapy. Methods of Gene therapy. Gene targeting and silencing. Gene therapy in the treatment of cancer, SCID, muscular dystrophy, respiratory disease (emphysema), cystic fibrosis. Challenges & future of gene therapy.

Course outcomes:

At the end of the course the student will be able to:

• Explain & compare the different vectors & enzymes used in the construction of recombinant DNA in Genetic engineering
• Choose& explain specific techniques like PCR, Blotting & construction of libraries
• Differentiate between & learn the different gene/DNA transfer techniques
• Outline the various methods of producing transgenic organisms and sub-divide/summarize the applications of genetic engineering for the welfare of mankind & society

Question paper pattern:

• The question paper will have ten full questions carrying equal marks.
• Each full question will be for 20 marks.
• There will be two full questions (with a maximum of four sub- questions) from each module.
• Each full question will have sub- question covering all the topics under a module.
• The students will have to answer five full questions, selecting one full question from each module.

Textbook/s

1 Principles of Gene Manipulation and Genomics S.B. Primrose and R. M. Twyman Blackwell Science Publications 7th edition 2006

2 Gene Cloning and DNA Analysis: An Introduction T A Brown Wiley – Blackwell Publications. (6th edition)

Reference Books

3 Recombinant DNA Watson .J.D. et al Scientific American Books, New York 1993

4 Plant Genetic Engineering J. H. Dodds Cambridge University Press 1983

5 Gene Cloning and Manipulation Howe C. J Cambridge University Press 2007