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Sathyabama Institute of Science and Technology B.Sc - Biotechnology SBB2102 Genetic Engineering Syllabus


SATHYABAMA INSTITUTE OF SCIENCE AND TECHNOLOGY SCHOOL OF BIO & CHEMICAL ENGINEERING

SBB2102 GENETIC ENGINEERING
L T P Credits Total Marks
3 0 0 4 100

UNIT 1 rDNA TECHNOLOGY AND TOOLS INVOLVED IN GENETIC MANIPULATIONS 12 Hrs.
Introduction to rDNA technology- pros and cons of genetic engineering. Restriction modification system,
Restriction enzymes – function, classification (Based on recognition and restriction sequence:-type I, II and III; based
on buffer salt concentration: - low, medium and high; based on pattern of restriction:-sticky (5’ and 3’) and blunt end
cutters). Other DNA modifying enzymes and its functions/uses in r-DNA technology (DNA Polymerases, Klenow
fragment, Ligase, S1 Nuclease, Mung Bean nuclease, Alkaline Phosphatase, Terminal Transferase, Polynucleotide
kinases, alkaline Phosphatases (CIP,SAP and TAP), RNase A, RNase H, DNase 1, Exonucleases, Reverse
Transcriptase)

UNIT 2 BIOLOGY OF CLONING VECTORS 12 Hrs.
Ideal features of vectors; Plasmids (Types, copy number, properties, origin of replication and incompatibility group,
plasmid amplification), bacteriophages eg λ (Life cycle, genome organization, feasibility as a cloning vehicle), Types of
Cloning Vectors (structure and general features of General Purpose cloning vectors, shuttle vectors), Examples of
cloning vectors (pBR322, pUC series of vectors, λ insertional and replacement vectors), derivatives of phages and
plasmids (cosmids, phagemids, phasmids) cloning vectors for large DNA fragments using YACs, PACs and BACs

Unit-3 INTRODUCTION TO R-DNA TECHNOLOGY 12 Hrs.
General strategies for isolation of genomic and plasmid DNA; Strategies for isolation of gene of interest (restriction
digestion, PCR), Creation of r-DNA (Restriction Digestion, modification of vector and insert, linker, adaptors,
homopolymer tailing, ligation,), PCR Cloning, Selectable and screenable markers, reporter genes.

UNIT 4 GENE TRANSFER TECHNIQUES 12 HRS.
Selection of host and vector, Host Organisms and its genotypes- Prokaryotic and eukaryotic systems, Methods of
gene transfer- Physical (micro injection, gene gun/biolistic, electroporation), Chemical (Calcium chloride, Calcium
phosphate precipitation method, liposome mediated) and Biological methods (Agrobacterium and viral).

UNIT 5 IDENTIFICATION OF GENETIC TRANSFORMANTS AND THEIR APPLICATIONS 12 Hrs.
Methods for clone identification-direct screening (insertional inactivation of marker gene, visual screeningmethods),
indirect screening (PCR and hybridization based techniques-colony PCR/ hybridization and dot blot hybridization),
hybridization techniques - Southern blotting, Northern blotting, Western blotting. Examples of Transgenic plants and
animals, current status of commercial rDNA products, Bio-safety measures and regulations for rDNA work

TEXT / REFERENCE BOOKS
1. Primrose, S.B. and Twyman, R.M., Principles of Gene Manipulation and Genomics, Blackwell Publishing
(2006) 7th ed. ISBN 1-4051-3544-1
2. Sambrook, Joseph and David W. Russell “ The Condensed Protocols: From Molecular
Cloning: A Laboratory Manual” Cold Spring Harbor , 2006.
3. Brown T.A., Genomes, 3 by Third Edition (Garland Science Publishing), 2007.
4. Glick , B.R. and J.J. Pasternak. “Molecular Biotechnology: Principles and Applications of Recombinant DNA”
4th Edition. ASM, 2010.

END SEMESTER EXAMINATION QUESTION PAPER PATTERN
Max. Marks : 100 Exam Duration : 3 Hrs.
PART A : 10 questions of 2 marks each - No choice 20 Marks
PART B : 2 questions from each unit of internal choice; each carrying 16 marks 80 Marks