SEMESTER – III
DSC – 2C
APMB 232 RECOMBINANT DNA TECHNOLOGY (3 credits)
Unit - I
Introduction to recombinant DNA technology – tools for rDNA technology – DNA manipulative enzymes: restriction enzymes, ligases, polynucleotide kinase, phosphatase, cutting of DNA molecules – joining of DNA molecules – homopolymer tails, linkers, adapters.
‘Unit - II
Gene cloning vectors: salient features, plasmids – properties, types, pBR322 and pUC18, bacteriophage vectors – l, lZAP, lgt11, cosmids, artificial chromosomes – BAC, YAC, MAC. Cloning bovine somatostatin gene in E. coli.
Unit - III
Transformation of r-DNA into target host organisms: calcium chloride mediated gene transfer, Agrobacterium mediated DNA transfer, electroporation, microinjection, liposome fusion, particle gun bombardment. Screening and selection of recombinant host cells: blue/white screening.
Unit - IV
Construction of gene libraries: genomic and cDNA libraries. Blotting techniques, polymerase chain reaction (PCR) and its applications.
Unit - V
Applications of rDNA technology in industry, medicine, agriculture and pharmacy. Social impact of recombinant DNA technology.
Text Books
1. Primrose S.B. and Twyman R.M. (2006) Principle of Gene Manipulation and Genomics,7th Edn. Blackwell Publishing.
2. Nicholl D.S.T (2008) An Introduction to Genetic Engineering, 3rd Edn. Cambridge University Press.
Reference Books
1. Brown T.A. (2016) Gene Cloning and DNA Analysis – An Introduction, 7th Edn. Wiley- Blackwell.
2. Glick B.R. and Patten C.L. (2017) Molecular Biotechnology. Principles and Application of Recombinant DNA, 5th Edn. ASM Press, Washington.
3. Watson J.D. Gann A., Baker T.A., Levine M., Bell S.P. and Losick R. (2014) Molecular Biology of Gene, 7th Edn. Pearson publishers.