Unit-I
Molecular tools and applications- restriction enzymes, ligases, polymerases, alkaline phosphatase. Gene Recombination and Gene transfer: Transformation, Episomes, Plasmids and other cloning vectors (Bacteriophage-derived vectors, artificial chromosomes), Microinjection, electroporation, Ultrasonication, Principle and applications of Polymerase chain reaction (PCR), primer-design, and RT- (Reverse transcription) PCR.
Unit-II
Restriction and modification system, restriction mapping. Southern and Northern hybridization. Preparation and comparison of Genomic and cDNA library, screening of recombinants, reverse transcription, Genome mapping, DNA fingerprinting, Applications of Genetic Engineering. Genetic engineering in animals: Production and applications of transgenic mice, role of ES cells in gene targeting in mice, Therapeutic products produced by genetic engineering-blood proteins, human hormones, immune modulators and vaccines.
Unit-III
Random and site-directed mutagenesis: Primer extension and PCR-based methods of site-directed mutagenesis, Random mutagenesis, Gene shuffling, production of chimeric proteins, Protein engineering concepts and examples.
Unit-IV
Genetic engineering in plants: Use of Agrobacterium tumefaciens and A. rhizogenes, Ti plasmids, Strategies for gene transfer to plant cells, Direct DNA transfer to plants, Gene targeting in plants, Use of plant viruses as episomal expression vectors.