GENETIC ENGINEERING AND BIOTECHNOLOGY 4(3-1)

Goal: To familiarize with the genetic engineering and biotechnology

Objectives:
• To develop skills of the students in understanding the basics of Genetic engineering
• To illustrate creative use of modern tools and techniques for manipulation and analysis of
genomic sequences
• To expose students to application of recombinant DNA technology in different branches
of Biotechnology

Course Contents:
The basic principles of Gene Cloning and DNA analysis. Vehicles for gene cloning. Properties of
prokaryotic and eukaryotic cloning vectors. Purification of DNA from living cells, Manipulation
of purified DNA: Nucleic acid modifying enzymes, Restriction endonucleases. Cloning strategies;
Introduction of DNA into living cells. Selectable and screening markers. Construction of genomic
and cDNA libraries, Study of gene location and structure: Probe labelling and hybridization,
Restriction mapping, Expression systems, DNA and RNA fingerprinting. Applications of
recombinant DNA technology in Agriculture sector, Applications of recombinant DNA
technology in animal sector, Impact of gene cloning in medicine, Site directed mutagenesis.
Practical: Methods of nucleic acid isolation (DNA & RNA), Electrophoresis, DNA amplification,
Gene cloning, Growing and harvesting a bacterial culture, Plasmid and genomic DNA isolation,
Restriction Digestion, Competent cells preparation, Ligation, Transformation, Screening of
transformed cells, Vector construction, Use of Bioinformatics software’s.
DETAILED COURSE OUTLINE:
No. Theory
1 Introduction to the course; sharing of learning outcomes
2 Gene Cloning
3 Vehicles for gene cloning
Structural differences between cloning and expression vectors
4 Structural and functional properties of Plasmids
5 Structural and functional properties of Bacteriophages
6 Histological features of avian bones and cartilages
7 Synthetic vectors
8 Purification of DNA from living cells
9 Purification of DNA from plant cells
10 Purification of extra chromosomal DNA from living cells
11 Manipulation of purified DNA
12 Restriction enzymes
13 Restriction digestion and mapping
14 Results detection using agarose gel electrophoresis
15 Results detection using autoradiography
16 Introduction of DNA into living cells (Transformation)
17 Competent cells preparation
18 Introduction of DNA into living cells (Transduction, Biolistic method, Transfection)
19 Introduction of DNA into living cells (Sperm mediated gene transfer)
20 Identification of recombinants (antibiotic resistance gene base, insertional
 inactivation)
21 Identification of transformed cells (Blue white screening)
22 Identification of transformed cells (Spi typing, Immunological screening)
23 Prokaryotic expression systems
24 Eukaryotic expression system (Pichia Pastoris)
25 Eukaryotic expression system (Pichia Pastoris)
26 Construction of genomic libraries
27 Differences between genomic and cDNA libraries
28 Mapping of genes
29 Probe labelling and hybridization
30 Control of gene expression
31 DNA and RNA fingerprinting
32 Brief introduction of Biotechnology
33 History of Biotechnology
34 Types of Biotechnology
35 Applications of recombinant DNA technology in Agriculture Biotechnology
(Anti sense technology)
36 Applications of recombinant DNA technology in Agriculture Biotechnology
(Transgenic crops)
37 Applications of recombinant DNA technology in Agriculture Biotechnology
(Transgenic crops)
38 Applications of recombinant DNA technology in Agriculture Biotechnology
(Transgenic animals)
40 Applications of recombinant DNA technology in Agriculture Biotechnology
(Transgenic animals)
41 Impact of Gene Cloning in medicine (Recombinant therapeutic proteins production)
42 Impact of Gene Cloning in medicine (Recombinant therapeutic proteins production)
43 Impact of Gene Cloning in medicine (Gene therapy)
44 Impact of Gene Cloning in medicine (Gene therapy)
45 Transpharmers - bioreactors for pharmaceutical products
46 Transgenic sheep for production of alpha Antitrypsin
47 Transgenic and gene knockouts
48 Transgenic and gene knockouts
No. Practical
1. DNA isolation from blood
2. RNA isolation from blood and tissues
3. DNA and RNA detection through gel electrophoresis
4. DNA and RNA quantification through Nano drop
5. Growing and harvesting a bacterial culture
6. Genomic and plasmid isolation from bacterial cells
7. Quantification of cells
8. Restriction digestion
9. Restriction digestion
10. Competent cells preparation
11. Gene ligation
 12. Blue white screening
13. Blue white screening
14. Vector construction
15. Vector constriction
16. Use of Bioinformatics software’s in gene cloning
Recommended Books/Readings:
1. Sambrook & Russel (2001). Molecular Cloning: A laboratory Manual. 3rd edition, Volume 1-
3, Cold Spring Harbor Laboratory press, New York, USA.
2. AlBerts B, Bray D, Lewis J, Raff M, Roberts K & Watson J. D (2002). Molecular Biology of
the cell. 4th edition.
3. Brown T. A (2001). Molecular Biology Lab. FEX. 2nd edition. Academic press Publication.
4. Richard Twyman (2014). Gene Transfer to Animal Cells. Garland Science
5. Primrose SB & Twyman (2013), "Principles of Gene Manipulation, An Introduction To
Genetic Engineering ", Blackwell Science Publication.
6. Sandhya Mitra (2008). Genetic Engineering Principles and Practice, Rajiv Beri for
Macmillan India Ltd publications.