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Home > GENETIC ENGINEERING

GENETIC ENGINEERING [1]

Paper Code: 
MBL 221
Credits: 
3
Contact Hours: 
45
Objective: 

Course Outcomes (COs):

 

Course Outcomes

Learning and teaching strategies

Assessment Strategies

 
 

Upon completion of the course the learner will:

CO 50: Acquaint and discuss tools for genetic engineering

CO 51: Apply various types of vectors in research.

CO 52: Explain and analyze types and applications of PCR techniques.

CO 53: Differentiate, compare and appraise Gene manipulation and protein-DNA interaction.

CO 54: Design mechanism and strategies of Gene silencing and genome editing technologies.            .

Interactive Lectures, Discussion, Tutorials, Reading assignments, Self-learning assignments, Effective questions, Giving tasks

Class test, Semester end examinations, Quiz, Solving problems in tutorials, Assignments, Presentation, Individual and group projects

 

 

11.00
Unit I: 
Introduction and tools for genetic engineering

Impact of genetic engineering in modern society; general requirements for performing a genetic engineering experiment; restriction endonucleases and methylases; DNA ligase, Klenow enzyme, T4 DNA polymerase, polynucleotide kinase, alkaline phosphatase; cohesive and blunt end ligation; linkers; adaptors; homopolymeric tailing; labelling of DNA: nick translation, random priming, radioactive and non-radioactive probes, hybridization techniques: northern, southern, south-western and far-western and colony hybridization, fluorescence in situ hybridization.

 

8.00
Unit II: 
Different types of vectors

Plasmids; Bacteriophages; M13 mp vectors; PUC19 and Bluescript vectors, hagemids; Lambda vectors; Insertion and Replacement vectors; Cosmids; Artificial chromosome vectors (YACs; BACs); Principles for maximizing gene expression expression vectors; pMal; GST; pET-based vectors; Protein purification; His-tag; GST-tag; MBP-tag etc.; Intein-based vectors; Inclusion bodies; methodologies to reduce formation of inclusion bodies; mammalian expression and replicating vectors Baculovirus and Pichia vectors system, plant based vectors, Ti and Ri as vectors, yeast vectors, shuttle vectors..

 

8.00
Unit III: 
Different types of PCR techniques

Principles of PCR: primer design; fidelity of thermostable enzymes; DNA polymerases; types of PCR – multiplex, nested; reverse-transcription PCR, real time PCR, touchdown PCR, hot start PCR, colony PCR, asymmetric PCR, cloning of PCR products; T-vectors; proof reading enzymes; PCR based site specific mutagenesis; PCR in molecular diagnostics; viral and bacterial detection; sequencing methods; enzymatic DNA sequencing; chemical sequencing of DNA; automated DNA sequencing; RNA sequencing; chemical synthesis of oligonucleotides; mutation detection: SSCP, DGGE, RFLP.

 

8.00
Unit IV: 
Gene manipulation and protein-DNA interaction

Insertion of foreign DNA into host cells; transformation, electroporation, transfection; construction of libraries; isolation of mRNA and total RNA; reverse transcriptase and cDNA synthesis; cDNA and genomic libraries; construction of microarrays – genomic arrays, cDNA arrays and oligo arrays; study of protein-DNA interactions: electrophoretic mobility shift assay; DNase footprinting; methyl interference assay, chromatin immunoprecipitation; protein-protein interactions using yeast two-hybrid system; phage display.

 

 

10.00
Unit V: 
Gene silencing and genome editing technologies

Gene silencing techniques; introduction to siRNA; siRNA technology; Micro RNA; construction of siRNA vectors; principle and application of gene silencing; gene knockouts and gene therapy; creation of transgenic plants; debate over GM crops; introduction to methods of genetic manipulation in different model systems e.g. fruit flies (Drosophila), worms (C. elegans), frogs (Xenopus), fish (zebra fish) and chick; Transgenics - gene replacement; gene targeting; creation of transgenic and knock-out mice; disease model; introduction to genome editing by CRISPR-CAS with specific emphasis on Chinese and American clinical trials.

 

ESSENTIAL READINGS: 
  • Brown, T. A., Gene Cloning and DNA Analysis, An Introduction.  Wiley –Blackwell publication. 2010.
  • Sandy B. Primrose, Richard M. Twyman.  Principles of Gene Manipulation and Genomics, Blackwell Sciectific Publication. 2009.
  • Brown, T. A., Genomes (3rd ed.). New York: Garland Science Pub. 2006.
  • Old, R. W., Primrose, S. B., & Twyman, R. M., Principles of Gene Manipulation: an Introduction to Genetic Engineering. Oxford: Blackwell Scientific Publications. 2001.
  • Green, M. R., & Sambrook, J., Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press. 2012.
  • Selected papers from scientific journals, particularly Nature & Science.

 

 

REFERENCES: 

SUGGESTED READINGS:

  • P.B. Kaufman, W. Wu. D. Kim and L.J; Cseke, Molecular and Cellular Methods in Biology and Medicine, CRC Press, Florida.
  • M. Glover and B.D. Hames, DNA Cloning: a Practical Approach, IRL Press, Oxford.
  • S.L. Berger and A.R. Kimmel, Methods in Enzymology vol. 152, Guide to Molecular Cloning Techniques, Academic Press, Inc. San Diego.
  • D.V. Goeddel, Methods in Enzymology Vol 185, Gene Expression Technology, Academic Press, Inc., San Diego.
  • Kingsman S.M. and Kingsman A.J., Genetic Engineering. An Introduction to gene analysis and exploitation in eukaryotes. Blackwell Scientific Publications, Oxford, 1990.
  • Desmond S.T. Nicholl, An Introduction to Genetic Engineering. Edited by, Cambridge University Press. 2002.
  • Sue Carson and Dominique Robertson, Manipulation and Expression of Recombinant DNA., Second edition, Academic Press. 2005.
  • Primrose and Twyman, Principles of Gene Manipulation and Genomics. (7th edition). Blackwell Publishers. 2006.

e RESOURCES:

  • https://nptel.ac.in/courses/102103013 [2]
  • https://nptel.ac.in/courses/102103074 [3]
  • https://www.youtube.com/watch?v=3IsQ92KiBwM [4]
  • https://www.youtube.com/watch?v=FtNreXY7poA [5]
  • https://www.youtube.com/watch?v=FvHRio1yyhQ&t=71s [6]

 

JOURNALS:

  • International Journal of Molecular Biotechnology
  • Indian Journal of Biotechnology
  • Current Science

 

 

 

 

Academic Year: 
2023-2024 [7]

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Source URL: https://biotech.iisuniv.ac.in/courses/subjects/genetic-engineering-24

Links:
[1] https://biotech.iisuniv.ac.in/courses/subjects/genetic-engineering-24
[2] https://nptel.ac.in/courses/102103013
[3] https://nptel.ac.in/courses/102103074
[4] https://www.youtube.com/watch?v=3IsQ92KiBwM
[5] https://www.youtube.com/watch?v=FtNreXY7poA
[6] https://www.youtube.com/watch?v=FvHRio1yyhQ&t=71s
[7] https://biotech.iisuniv.ac.in/academic-year/2023-2024