MTI - 505 GENETIC ENGINEERING - I

UNIT 1: Introduction to Genetic Engineering
Scope and history, Handling of DNA, RNA Restriction Enzymes, radioactive
isotopes and other corrosive, carcinogenic and inflammable chemicals/ reagents

UNIT 2: Requirement of genetic engineering
Isolation and purification of nucleic acids from living cells. DNA isolation:
Preparation of total cell DNA from animal and plant, Preparation of Plasmid
DNA, preparation of Bacteriophage DNA. RNA isolation: Total Cellular RNA,
Cytoplasmic and NuclearRNA, Poly ( A+) RNA

UNIT 3: Enzymes in recombinant DNA technology
Nucleases, Ligases, Poylmerases, DNA modifying enzymes, Topoisomerases,
Restriction endonucleases

UNIT 4: Cloning Vectors:
Nomenclature, properties of good vector. Vectors for E. coli: Plasmid and plasmid
based vectors: BAC, Phagemids, cosmids and expression, Virus based vectors:
M-13, Lambda and PAC. Vectors for eukaryotes: Yeast based vectors : 2micro m
plasmid, YEp, YAC, Vectors for plants : Ti plasmid and Ri Plasmid Ca MV,
vectors for Animals - SV-40, Retrovirus vectors and Vaccinia/ Bacculo virus

UNIT 5: In Vitro construction of recombinant DNA molecules
Creation of r-DNA, Gene Libraries – Genomic DNA and c DNA Cloning
techiniques, Selection of clones based on antibiotic, insertional inactivation,
hybridization, PCR and antibody.

UNIT 6: Methods of DNA transformation in bacteria, plants and animal cells
Salt based method, Electroporation, Genegun, Liposome mediated, Transfection