Pre-requisites : Nil
The basis of genetic engineering: Genome organization; the flow of genetic information; Genes and genomes; Genome size and complexity; The transcriptome and proteome. Labelling nucleic acids; End labelling; Nick translation; Labelling by primer extension; Nucleic acid hybridization. DNA sequencing; Principles of DNA sequencing; Next-Gen sequencing. Human Genome Project.
Techniques in Genetic Engineering: DNA/ RNA extraction, Polymerase Chain reaction: primer design, Nested PCR, inverse PCR, RAPD, Real-time PCR. Gel Electrophoresis-DNA, RNA and Protein. Blotting: DNA, RNA and Protein. Site directed mutagenesis, DNA/ Protein interactions: CHIP, EMSA, Reporter assays, pull down assays.
DNA modifying enzymes and vectors: Nucleases; Polymerases; Enzymes that modify the ends of DNA molecules: Terminal Transferase, T4 Polynucleotide Kinase, Alkaline Phosphatases; DNA ligase, Restriction enzymes. Vectors for cloning and expression in prokaryotes and Eukaryotes: Plasmid Vectors, Phage Vectors, Cosmids, Phagemids, BACs, Yeast
Vectors, YACs, Lentiviral Vectors, Adenoviral Vectors, Plant Vectors, Insect Vectors. In-frame protein fusion and use of GFP for subcellular localization of protein/DNA. cDNA and genomic DNA library construction. Introduction of foreign DNA: Transfection, transformation, particle gun and Ti plasmid mediated.
Recent advances and applications: ZFNs, TALENs, CRISPR/Cas systems. Transgenic plants and animals, gene therapy, forensic applications.
Bioinformatics: DNA and protein databases. Promoter identification, Gene expression analysis, Big data analysis.
1. Desmond S. T. Nicholl, An Introduction to Genetic Engineering (3rd Ed), Cambridge University Press, 2010.
2. T. A. Brown, Gene Cloning and DNA Analysis: An Introduction, (6th Ed), Wiley-Blackwell, 2012.
3. Sandy B. Primrose, Richard Twyman, Principles of Gene Manipulation and Genomics, (8th Ed), Blackwell Publishing, 2016
4. Michael R. Green and Joseph Sambrook, Molecular Cloning: A Laboratory Manual, (4th Ed), Cold Spring Harbor Laboratory Press, 2012.
1. Bernard R. Glick, Jack J. Pasternak, Cheryl L. Patten, Molecular Biotechnology: Principles and Applications of Recombinant DNA (4th Ed), ASM Press, 2010.
2. T.A. Brown, Introduction to Genetics: A Molecular Approach, (1st Ed), Garland Science, 2011.
3. Jordanka Zlatanova, Kensal van Holde, Molecular Biology, (1st Ed), 201