BT8601 - GENETIC ENGINEERING (Syllabus) 2017-regulation Anna University

BT8601 - GENETIC ENGINEERING (Syllabus) 2017-regulation Anna University

BT8601	GENETIC ENGINEERING	LPTC

3003

OBJECTIVES:

• To discuss the gene cloning methods and the tools and techniques involved in gene cloning and genome analysis and genomics.
• To explain the heterologous expression of cloned genes in different hosts.

UNIT I	BASICS OF RECOMBINANT DNA TECHNOLOGY	12

Manipulation of DNA – Restriction and Modification enzymes, Design of linkers and adaptors. Characteristics of cloning and expression vectors based on plasmid and bacteriophage, Vectors for insect, yeast and mammalian system, Prokaryotic and eukaryotic host systems, Introduction of recombinant DNA in to host cells and selection methods.

UNIT II	DNA LIBRARIES	12

Construction of genomic and cDNA libraries, Artificial chromosomes – BACs and YACs, Chromosomal walking, Screening of DNA libraries using nucleic acid probes and antisera.

UNIT III	SEQUENCING AND AMPLIFICATION OF DNA	12

Maxam Gilbert’s and Sanger’s methods of DNA sequencing. Inverse PCR, Nested PCR, AFLP- PCR, Allele specific PCR, Assembly PCR, Asymmetric PCR, Hot start PCR, inverse PCR, Colony PCR, single cell PCR, Real-time PCR/qPCR – SYBR green assay, Taqman assay, Molecular beacons. Site directed mutagenesis.

UNIT IV	ORGANIZATION AND STRUCTURE OF GENOMES	12

Organization and structure of genomes, Genome sequencing methods, Conventional and shotgun genome sequencing methods, Next generation sequencing technologies , Ordering the genome sequence, Genetic maps and Physical maps, STS content based mapping, Restriction Enzyme Finger Printing, Hybridization mapping, Radiation Hybrid Maps, Optical mapping. ORF finding and functional annotation.

UNIT V	CURRENT STATUS OF GENOME SEQUENCING PROJECTS	12

Current status of genome sequencing projects, Introduction to Functional genomics, Microarrays, Serial Analysis of Gene expression (SAGE), Subtractive hybridization, DIGE, TOGA, Yeast Two hybrid System, Comparative Genomics, Proteogenomics, Web resources for Genomics, Applications of genome analysis and genomics.

TOTAL : 60 PERIODS

OUTCOMES:

• The students after completing this course would be aware of how to clone commercially important genes.
• The students would be aware of how to produce the commercially important recombinant proteins.
• The students would be aware of gene and genome sequencing techniques.
• The students would be aware of microarrays, Analysis of Gene expression and proteomics.

TEXT BOOKS:

1. Old RW, Primrose SB, “Principles Of Gene Manipulation, An Introduction To Genetic Engineering “, Blackwell Science Publications, 1993.
2. Principles of Genome Analysis and Genomics by S.B.Primrose and R.M.Twyman, 3rd Ed. (Blackwell Publishing)

REFERENCES:

1. Ansubel FM, Brent R, Kingston RE, Moore DD, “Current Protocols In Molecular Biology“Greene Publishing Associates, NY, 1988.
2. Berger Sl, Kimmer AR, “Methods In Enzymology”, Vol 152, Academic Pres