Elective III Course No.: BT-4XX S.

No 1 2 3 L T P: 3 0 0 Credits: 3 LT P 300 300 300 Credits 3 3 3

Course No Name of course BT-420 Recombinant DNA Technology & Application BT-421 BT-422 Animal Cell Culture Technology Molecular Pathogenesis Recombinant DNA Technology & Applications

Course No.: BT- 420 L T P: 3 0 0 Credits: 3 Objective: This course is designed to impart the knowledge of modern techniques used in genetic manipulations leading to the creation of genetically modified organisms, plants etc. Course Outline: Tools of Genetic Engineering: Cloning Vehicles, Restriction Enzymes, Modifying Enzymes, DNA Ligase, Polymerase Etc. Cloning Vectors: Plasmids, Lambda Phage, Phagemids, Cosmids, Artificial Chromosomes (Bacs, Yacs), Shuttle Vectors, Virus Based Vectors. Methods Of Gene Transfer: Transformation, Transduction, Particle Gun, Electroporation, Liposome Mediated, Microinjection, Agrobacterium Mediated Gene Transfer. Preparation and Application Of Molecular Probes: DNA Probes, RNA Probes, Radioactive Labeling, Non Radioactive Labeling, Use Of Molecular Probes, DNA Fingerprinting. Analysis and Expression of Cloned Gene: in Host Cells: Expression Vectors, Restriction Enzyme Analysis, Southern Blotting, Northern Blotting, Western Blotting, InSitu Hybridization. Colony and Plaque Hybridization, Factors Affecting Expression of Cloned Genes, Reporter Genes, Fusion Proteins. Gene Libraries - DNA Synthesis, Genomic DNA Libraries, Amplification of Gene Libraries, Identifying the Products Of cDNA Clones. Isolation, Sequencing and Synthesis of Gene: Different Methods of Gene Isolation, Techniques of DNA Sequencing, Artificial DNA Synthesis. Polymerase Chain Reaction (PCR): Basic Principles, Modifications, Applications. Modifying Genes: Site-Directed Mutagenesis, Insertion & Deletion Mutagenesis. Text / Reference Books: 1. From Genes to Clones by Winnacker. Panima 2. Molecular Biotechnology by Pasternack and Glick. 3. from Genes to Genomes: Concepts & Applications of DNA Technology by J.W. Dale & M.V. Schartz. 4. Gene Cloning & DNA Analysis: An Introduction (4th Edition) By T.A. Brown. 5. Molecular Cloning By Sambrook, Et Al. 6. Principles of Gene Cloning By Old and Primrose.

Animal Cell Culture Technology Course No.: BT-421 L T P: 3 0 0 Credits: 3 Objective: This course is designed to impart the knowledge of structure and organization of animal cells, medium requirement and control parameters used in the maintenance and production of Animal cell cultures, stem cell culture and the technology used for its storage and applications. Course Outline: Structure and Organization of Animal Cell: Equipments and materials for animal cell culture technology; Primary and established cell line cultures; Introduction to the balanced salt solutions and simple growth medium. Cultural and Nutritional Requirement: Brief discussion on the chemical, physical and metabolic functions of different constituents of culture medium. Role of carbon dioxide. Role of serum and supplements; Serum & protein free defined media and their application. Growth Characteristics: Basic techniques of mammalian cell culture in vitro; disaggregation of tissue and primary culture, maintenance of cell culture; cell separation. Measurement of viability and cytotoxicity; Biology and characterization of the cultured cells, measuring parameters of growth. Cell Culture: Cell synchronization; Cell cloning and micromanipulation; Cell transformation; Application of animal cell culture; Scaling-up of animal cell culture. Stem Cell Cultures: embryonic stem cells and their applications; Cell culture based vaccines, Somatic cell genetics. Organ and Histotypic Cultures: Measurement of cell death; Apoptosis; Three dimensional culture and tissue engineering. Text /References Books 1. Culture of Animal Cells, (3rd Edition), F1. Ian Froshney. Wiley-Liss. 2. Animal Cell Culture Practical Approach, Ed. John R.W. Masters, OXFORD. 3. Cell Culture Lab Fax. Eds. M. Butler & M. Dawson, Bios Scientific Publications Ltd., Oxford. 4. Animal Cell Culture Techniques. Ed. Martin Clynes, Springer. 5. Methods in Cell Biology, Vol. 57, Animal Cell Culture Methods. Ed. Jenni P. Mather and David Barnes. Academic Press. 6. Cell Growth and Division: A Practical Approach. Ed. R. Basega, IRL Press.

Molecular Pathogenesis Course No.: BT-422 L T P: 3 0 0 Credits: 3 Objective: This course is designed to impart the knowledge of pathogens and their growth and behavioral patterns, characteristics for the diagnosis, host defense mechanism against pathogens and solutions for pathogen caused diseases. Course Outline: Introduction: Introduction to pathogenesis, components of microbial pathogenicity. Population genetics of Microbial pathogenesis, methods to detect genetic diversity and structure in nature population, epidemiology, cryptic diseases. Host Defense I: Host defense against pathogens, clinical importance of understanding host defense, components of the host surface defences systems like skin, mucosa, eye, mouth, respiratory tract. Host Defense II: Components of the systemic defense like the tissues and blood. Host- Pathogen Interaction: Virulence and virulence factors, colonising virulence factors, virulence factors damaging the host tissues, virulence genes and regulation of the virulence genes. Experimental Methods: Experimental methods to study host-pathogen interaction, selecting the pathogen model, measurement of virulence, identification of potential virulence factors. Modulation of Immune Response: modulation of immune response by vaccines, properties of vaccines, other immuno modulators. Paradigms of Pathogenesis: Diphtheria disease by colonisation; Disease without colonisation, Clostridium botulinum and Staphylococcus aureus; Intestinal infections, Shigella and E.coli infections; Vibrio cholera Salmonella infections; fungal infections Future Challenges: Gastric and duodenal ulcers - are they due to infections? Lyme disease and Syphilis - unsolved mystery. Legionnaires disease-aftermath of comforts.Tuberculosis and other mycobacterial infections reemerging with vengeance. Rheumatic fever and glomerulo nephritis - still a question to be solved.

Text / References Books: 1. Iglewski B.H. and Clark V.L. Molecular basis of Bacterial pathogenesis, Academic press, 1990. 2. Janeway C.A. Jr, and Travers P. T. Immunobiology. Blackwell J Scientific Publishers, 1994. 3. Talaro K. and Talaro A. Foundations in Microbiology, W.C. Brown Publishers, 1993. 4. Roitt I. Essentials of Immunology, 8th edition, Blackwell Scientific Publishers, 1994. 5. Austyn J.M. and Wood K.J. Principles Cellular and Molecular Immunology, OxfordUniversityPress,1993.