UTechBiotech.org, Kolkata, India

Recombinant DNA Technology
 
L-T- P (3-0-0)
Total Credits 3
       
  Recommended Texts & References:  
    1) Principles of Gene Manipulation, Primrose et al
2) Recombinant DNA, Watson et al
3) Molecular Biotechnology, Glick & Pasternak
4) Molecular Cloning, Sambrook & Russel
5) Instant Notes, Molecular Biology, Turner et al
6) Molecular Biology, Robert Weaver
  
     
  Module I: Expression of genes in prokaryotic and eukaryotic systems
4 Lecture Hours
   

Use of prokaryotic and eukaryotic organisms as Biological systems in Molecular Biotechnology; Gene structure in prokaryotic and eukaryotic cells; Genetic elements that control Gene expression – concept of operon and related elements in the unit, regulatory and structural gene, post translational processing of mRNA, extra chromosomal DNA and its functions.

       
  Module II: Tools in rDNA technology
10 Lecture Hours
   

Restriction endonucleases and other enzymes used in recombinant DNA technology; Cloning vectors: Plasmid cloning vector pBR322, other plasmid vectors, Bacteriophage l vectors and other phage vectors; Cosmids, Phagemids; YAC and BAC vectors; Genetic transformation of prokaryotes: Transferring DNA into E.coli – Chemical induction and Electroporation; DNA labeling -radioactive and non-radioactive methods; DNA sequencing – Chemical cleavage and dideoxy methods; Polymerase Chain Reaction, RT-PCR, RACE, Real-time quantitative PCR; Southern and Northern blotting,  DNA fingerprinting
       
  Module III: Analysis of cloned genes and studying gene function
16 Lecture Hours
   
Construction of cDNA and genomic DNA library: library screening by DNA hybridization, immunological assay, and protein activity; gene isolation and cloning; Chromosome walking, in situ hybridization, site-directed mutagenesis, Expression of cloned gene in recombinant cells – production of biochemicals; DNA markers; Gene transfer to animal cells: strategies (direct transfer, transfection), Reporter genes and promoter analysis, Selectable markers; Two-vector expression system. Cloning in bacteria other than E. coli / Saccharomyces cerevisiae and other fungi.
  
       
  Module IV: Application of rDNA technology
12 Lecture Hours
   
DNA Diagonostic system: random amplified polymorphic DNA (RAPD), DNA finger printing, and their applications. Advances in Transgenic Technology; Antisense and ribozyme technology, Human genome project and its application, Gene therapy prospect and future, Current production of rDNA products, Bio-safety measures and regulations for rDNA work.
  
       
   
 
  

3 Credit (Theory) means 3 lecture hours each week or 42 lectures per semester.
2 Credit (Lab) means at least 40 hours of lab work per semester.