Syllabus of Jawaharlal Nehru University MSc Biotechnology - BT521 - Molecular Biology and Molecular Genetics

Credits: 5

1. What is Molecular Biology?
DNA as the hereditory material. RNA can also be the hereditory material, but only in some viruses.
Determination of radioactivity.

2. Size of natural DNA molecule.
Estimation of DNA
Difference between DNA and RNA. Presence of phosphodiester linkage in RNA and DNA. 5' to 3' linkage.
Double strandedness of cellular DNA.

3. Antiparallel nature of the two strands of DNA.

4. Denaturation and renaturation of DNA. Tm. GC content from Tm. Renaturation kinetics of DNA and complexity of DNA. Cot curves.

5. Separation of and determination of single stranded and double stranded DNA.
DNA-DNA hybridization-relatedness of difference genes and species.

6. Restriction endonuclease - an essential tool for gene cloning. DNA ligases. Plasmids. Antibiotic resistance markers.
Transformation of E. coli.

7. Desirable characteristics of a plasmid cloning vector.

8. Centrifugation - velocity and equilibrium.
Messelson & Stahl experiment.

9. Replication fork - John Cains experiment. Huberman & Riggs experiment on replication of Chinese Hamster DNA. Inman experiment on replication of lambda DNA.

10. Genetic mapping - recombination frequency in bacteria and viruses - conjugation - interrupted mating. Taylor map. Hfr strains.

11. Terminal redundancy and Circular permutation in T even phages.

12. DNA content in E. coli. Dichotomous replication.

13. Synchronization of E. coli cells.

14. Bi-directional replication in E. coli.

15. Mechanism of genome transfer during conjugation
Between Hfr and F strains of E. coli.

16. Mechanism of Genetic recombination.

17. Kornberg DNA polymerase.

18. Exonuclease II and exonuclease VI as integral part of Kornberg DNA polymerase.

19. Okazaki fragments.

20. The Pol AI mutant. DNA pol II and pol III. Kornberg's reappraisal of his old work. Properties of pol I, pol II and pol III. Phenotyphic defects of pol A mutant. Ts mutants of DNA replication and genetic mapping of the mutant loci.

21. Conversion of single stranded circular DNA viruses into double stranded RF form. Synthesis of viral plus strand from RF DNA.

22. OX gene A protein and its cleavage site. Rolling circle mode of synthesis. Involvement of SSB protein.

23. Overlapping genes.

24. Insertion sequences.
Transposons.

25. Genome organization of prokaryotes
Viral genomes : General information about different types of viral genomes. Structural organization of Retroviral genomes.

26. Transcription :

i. Flow of information from DNA to protein. Organization of genes in bacteria. Colinearity of genes and proteins. Operon concept.

ii. Process of transcription :
RNA polymerase subunit structure and function role of sigma factor in differential expression of genes in bacteria.

Transcription units and Cis elements. Promoter :
Consensus sequences affecting the promoter function. Constitutive and inducible promoters. Operator sequences as regulatory cis sequences.

Initiation : Interaction of polymerase with the promoter and control at initiation. Attenuation. Elongation. Termination : Rho dependent and Rho independent termination. Control at termination : Attnuation. Antitermination.

Processing of primary transcripts in prokarytoes :
Processing of tRNA and rRNA. Cleavage of T7 early mRNAs by RNase III. Control at the processing level.

iii. Regulation of transcription in bacteria : Introduction and repression. Repressor as a regulatory molecule. Coordinated control of gene clusters. Positive and negative regulation : Regulation of transcription of lac, trp, ara, his, and gal operons. Regulation through catabolite repression. CAP protein as a positive control factor.

iv. Transcriptional regulation in bacteriopohage Lambda. Lytic and Lysogenic switch. Role of various regulatory proteins.

v. Sequential expression of genes in Bacteriophage T4, 2 T7, Mu, x 174 : Genome organization and infection. Lectures

27. Translation :
Genetic code and codon bias. Essential components of translation. Ribosome : the site for translation,sub unit composition and assembly. Role of ribosomal RNA in translation. tRNA : Salient features of tRNA. Aminoacyl Trna synthetases. Difference between initiator fmett RNA and mettRNA, Suppressor tRNAs, frameshift suppression.Codon Anticodon recognition :Wobble hypothesis. Process of translation : Activation, Initiagtion, elongation translocation and termination. Factors involved in various steps.Peptidyltransferases.Transport of bacterial proteins : Cotranslational and Post translational mechanisms. Control of geneexpression at translational level.