Aims and Objective

The course introduces students to a study of the basic Principles of Molecular Biology. This course aims at introducing to students about:

DNA replication models, Transcription and Translation, Protein synthesis in the prokaryotes and eukaryotes, Processing and storing of proteins in the cell, Understanding of molecular Biology techniques for analysis of DNA, RNA, Proteins, regulation of gene expression, modern biophysical techniques including cell disruption and lysis methods, ultracentrifugations, electrophoresis, NMR and X-Ray Crystallography

Learning Outcome

This subject will integrate the concept and techniques developed in Molecular Biology, Physical Biology and biochemistry. Students will be expected to gain an understanding of the DNA, RNA, and protein synthesis and applications of biophysical techniques. Upon completion of this course students will be able to describe various biochemical and Biophysical techniques to characterise DNA, RNA and proteins.

Course Pre requisites 

General Biochemistry Course 

Syllabus

Molecular dogma; DNA as a genetic material. DNA replication in prokaryotes and Eukaryotes. Type of DNA polymerases and functions. DNA damage; types of mutations. DNA repair; NER, MMR, homologous DNA repair.

Virus DNA/RNA replication and its protein regulation.

Transcription in prokaryotes and Eukaryotes; differences and types of DNA polymerases, operons. Splicing; introns, exons and ribozymes. Gene regulation.

Translation; protein synthesis, regulation. Metabolism of RNA and DNA nucleosides. Structure of Chromatin and its functions.

Protein expression, purification and characterization using different biophysical methods i.e. UV/Vis Spectrophotometry, FT-IR, Circular Dichroism (CD), Surface Plasmon Resonance (SPR), Gel electrophoresis (SDS-PAGE, agarose gel electrophoresis and capillary electrophoresis), Ultra-, analytical and gradient centrifugation, Cryo-electron microscopy, protein NMR, X-ray crystallography/Diffraction. DNA amplification by PCR and real time PCR. Applications of DNA sequencing, Mass spectrometry and isotopes in molecular biology.

Assessment Criteria

Sessional (15) : Quizzes/Tests, Assignments, Presentations/ Attendance, Class Participation

Midterm (15): Subjective (short questions) and Objective Questions 

Final (45): Subjective (short and long questions) and Objective Questions 

Lab (25): Practical exam, Participation, Lab Book, Viva Exam

Time Table

Every Monday to Wednesday:  2:00 pm - 3:00 pm

Text Books and Reading Lists

1. Lehninger, A. L. Principles of Biochemistry, W. H. Freeman; Seventh edition (2017)

2. Voet, D. and J. G. Voet,. Fundamentals of Biochemistry: Life at the Molecular Level, Wiley; 5 edition (2016)

3. Zubay, G. Biochemistry, 4th Edition Macmillan Publishing Co.(1999).

4. Stryer, L. Biochemistry, Freeman & Co.( 1994)

WEEKLY LECTURE SCHEDULE

Labs -

1. Methods for the isolation of proteins from plant and animal samples i.e. liver/plasma protein, Salivary and Barley amylase using various cell disruption methods.
2. Protein precipitation by NH4SO4 method, using acid and organic solvent methods.
3. Protein dialysis and ultrafiltration methods.
4. Estimation of proteins using UV, Braford and Lowry’s methods.
5. Characterization of proteins i.e. amylase activity by enzyme assays, plasma proteins by characterization by SDS PAGE and Western blotting, analysis of a Protein From Egg White
6. Chromatographic separation/purification of proteins
7. Characterization of protein secondary structure using UV, FT-IR and circular dichroism spectroscopy.
8. Estimation and Isolation of total DNA/RNA from plant/animal tissues/cells, preparation of genomic/plasmid DNA from bacteria. Phenol/chloroform extraction of DNA. Mini- and Maxi- preparation of DNA
9. Isolation of RNA from mammalian cells
10. Characterization of DNA by Agarose Gel Electrophoresis and Southern blotting
11. Primer design and amplification of target DNA by PCR
12. Restriction enzyme digestion
13. Preparation of competent cells and gene cloning
14. Preparation of sequencing gels and DNA sequencing
15. PCR primer designing by online resources
16. Characterization of proteins using online tools.