Unit

Topic

No. of Lectures/Hrs

1

Recombinant DNA Technology: Enzymes for DNA manipulation, Restriction Endonucleases (Types I-IV, biological role and application); Restriction Mapping (Linear and Circular); Basic idea about CRISPR gene editing, DNA and RNA Purification, Vectors: Prokaryotic (pUC 18 and pUC19, pBR322, Ti plasmid, BAC); Lambda phage, M13 phagemid, Cosmid, Shuttle vector; Eukaryotic Vectors (YAC), Bacterial Transformation and selection of recombinant clones, Recombinant Screening

10

2

Introduction of DNA to host cells: Agrobacterium mediated gene transfer, Direct gene transfer by Electroporation, Microinjection, Microprojectile bombardment; Selection of transgenics– selectable marker and reporter genes (Luciferase, GUS, GFP)

8

3

Method of Clone identification, Expression and Modification: DNA Library, construction of genomic and cDNA libraries, screening DNA libraries to obtain gene of interest by genetic selection; complementation, colony hybridization, Expression Vector, DNA microarray technology, Site directed mutagenesis.

9

4

PCR and Reverse Transcriptase-PCR, Nucleic acid hybridization, Isotopic and non-Isotopic labelling of Nucleic acids, Blotting techniques: Northern, Southern and Western Blotting, DNA Fingerprinting; Molecular DNA markers i.e. RAPD, RFLP, SNPs; DNA sequencing.

9

5

Applications of Biotechnology: Pest resistant (Bt-cotton); herbicide resistant plants (Round Up Ready soybean); Transgenic crops with improved quality traits (Flavr Savr tomato, Golden rice); Improved horticultural varieties (Marigold); Role of transgenics in bioremediation (Superbug); edible vaccines; Industrial enzymes (Protease, Lipase); Genetically Engineered Products–Human Growth Hormone; Humulin; Biosafety concerns.

12

6

Plant Tissue Culture: Historical perspective; Composition of media; Nutrient and hormone requirements (role of vitamins and hormones); Totipotency; Organogenesis; Embryogenesis (somatic and zygotic); Protoplast isolation, culture and fusion; Tissue culture applications (micropropagation, androgenesis, virus elimination, secondary metabolite production, haploids, triploids and hybrids; Cryopreservation; Germplasm Conservation).

12