Post Graduate in Recombinant DNA Technology

This paper provides the student a thorough knowledge in principles and methods
in genetic engineering, vectors in gene cloning, transformation in higher organisms,
Human Genome Project and gene therapy
Objective: The main objective of the paper is to expose students to application of rDNA
technology various fields of biotechnology, medicine and research areas
Goal: This paper will help the student get a grasp of the latest advances in recombinant
DNA technology, which is a powerful tool in modern Biotechnology
UNIT- I
Principles and methods in genetic engineering: Isolation and purification of Nucleic
Acids - Agarose Gel Electrophoresis - Southern, Northern and South-Western blotting
techniques - Principles and techniques of nucleic acid hybridization and cot curves -
Polymerase Chain Reaction: Methods and advancements - Enzymes in Molcular
Biology: Nucleases, Restriction endonucleases, DNA Ligases, topoisomerases, gyrases,
methylases, other modifying enzymes - Bacterial Transformation: Principles and
methods.
UNIT-II
Vectors in Gene Cloning: Plasmids, Bacteriophages, Phagemids, Cosmids - Artificial
Chromosomes: PAC, BAC, YAC - Cloning in Prokaryotes E.coli - Cloning in Organisms
other than E.coli: Pseudomonas, Bacillus subtilis, Yeast and Fungi - Two and three
hybrid systems - Gene expression in Prokaryotes: merits and demerits - examples.
UNIT-III
Transformation in higher organisms: Plants - T-DNA and Transposon tagging -
Agrobacterium mediated transformations - Alternate transformation strategies:
chloroplast transformation etc. - Construction of Transgenic crop plants - Examples -
Markers for Transgenics. Animal transgenics: DNA mediated transformation - Gene
transfer by viral transduction - Viral vectors for Animal transgenics - Genetic
manipulation of mammals - DNA transfer to other vertebrates - - Transgenic and gene
knock out Technologies - Animal models for transgenic studies.
UNIT-IV
.Site-directed Mutagenesis - Fusion proteins: GFP and other examples. Genomic library -
cDNA library: Screening Representative cDNA libraries - RT PCR - RACE PCR, DDPCR.
Site directed Mutagenesis: Screening recombinant clones. Functional genomics:
Nucleic acid microarrays and Real Time PCR. RNAi and Gene Silencing. - Genome
analysis: DNA Sequencing and annotations: Methods and advancements - The Human
Genome Project.
UNIT-V
Gene Therapy: Different strategies for Gene Therapy, Si RNA, antisense RNA, Gene
therapy for inherited diseases, ADA, FH, Cystic Fibrosis, DMA, Neoplastic disorders,
infectious diseases., DNA forensics, , Somatic Cell Gene therapy, Triple helix
therapeutics, Antisense oligonucleotides, Ribozymes, Aptamers. Molecular Breeding of
Plants - Production of interferons - DNA vaccines - Gene targeting: Applications in
medicine, agriculture and industry