Biochem-703

BIOCHEM-703     RECOMBINANT DNA TECHNOLOGY AND GENE MANIPULATION                   3(2-1)

Learning objectives

By the end of this course students will be able to:

1. Have basic and recent knowledge about recombinant DNA technology
2. Explain genetic engineering and biotechnology
3. Perform different techniques of recombinant DNA technology
4. Know about basic bioinformatics tools

 Theory

Introduction to cloning strategies; Isolation of DNA and RNA from living material; Gene Isolation: Polymerase chain reaction (PCR) and RT-PCR; Introducing recombinant plasmids into suitable host: Tools, enzymes, vectors and hosts; Recombinant selection and characterization; RT-PCR. DNA sequencing; Sanger and Next Generation; Chemical synthesis of Primers and Genes; Probes: synthesis and detection. Cloning in mammalian and plant cells; DNA fingerprinting and its application in forensic sciences. Gene expression: Real-time PCR, reporter genes, fusion proteins, differential-display PCR, microarray, RNA sequencing, in vitro translation; Introduction to proteomics; Antisense RNA technology; CRISPER-CAS technology; Application and uses of genetic engineering in agricultural and biochemical research; Bioinformatics tools for the study of gene structure and function.

Practical

Preparation of solutions for molecular biology; Detection and quantitative determination of DNA; Cloning experiments using E. coli as host; Isolation and qualitative detection of plasmid DNA (miniprep.); Digestion of DNA with restriction enzymes and separation of different sized fragments on agarose gel; Study of transformed bacteria on the basis of antibiotic resistance; DNA amplification by polymerase chain reaction; PCR-RFLP.

 Suggested Readings

1. Alberts, B., A. Johnson, J. Lewis, D. Morgan, M. Raff, K. Roberts and P. Walter. 2014. Molecular Biology of the Cell. 6^th Garland Sciences. Taylor and Francis Group, New York, USA.
2. Brown, T. A. 2010. Gene Cloning and DNA Analysis: An Introduction, 6^th Wiley-Blackwell, UK.
3. Green M. R., and J. Sambrook. 2012. Molecular Cloning: A Laboratory Manual. 4^th Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA.
4. Lodish, H.,  Berk, C. A. Kaiser, M. Krieger andA. Bretscher. 2016. Molecular Cell Biology. 8^th Ed. Freeman, W. H. and Company. New York, USA.
5. Nelson, D.L and M.M. Cox. 2021. Lehninger Principles of Biochemistry. 8^th Worth Publishers, New York, USA
6. Weaver, R. F. 2012. Molecular Biology. 5^th Ed. McGraw Hill, New York, USA.

7. Ali, M. et al., Experimental Biochemistry and Biotechnology (Series I), 2019, 2^nd edition, ISBN: 978-969-23317-1-5.

8. Ali, M. et al., Experimental Biochemistry and Biotechnology (Series II), 2019. 1^st edition, ISBN: 978-969-23317-2-2.

9. Ali, M. et al., Experimental Biochemistry and Biotechnology (Series III), 2022, 1^st edition, ISBN: 978-969-23317-6-0.