| December 2nd, 2020 11:37 AM | |
| prince karak | Sathyabama Institute of Science and Technology B.Sc - Biotechnology SBB2102 Genetic Engineering Syllabus 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 |
