Cellular organelles – Structure and function. Organization of cytoskeleton and nuclei. Chromatin structure. Molecular aspects of cell division and cell cycle. Cell fusion. Nuclear – cytoplasmic interactions. Structure of DNA and RNA melting an dunwinding kinetics. DNA replication, kinetics, Repair and gene amplification. RNA transcription and processing. Protein synthesis and secretion. Transcriptional and Translational controls. Hormones and gene expression. Viruses and gene expression. Growth factors Mitogens and oncogenes. Cellular regulation, development and differentiation.

Enzyme classification and nomenclature. General properties of enzymes like effect of pH, temp, ions etc. Extraction , assay and purification of enzymes. Steady state kinetics. Michaelis – Menten, Lineweaver-Burke, Eadie-hofstee and Hanes-Woolf equations and Km value. Enzyme inhibitors. Pre-steady state kinetics. Fast kinetics to elucidate the intermediates and rate limiting steps (Flow and Relaxation methods). Enzyme specificity. Evidences for enzyme substrate complex. Nucleophilic attack. Role of metal ions in enzyme enzyme catalysis. Mechanism of enzyme action eg. Lysozyme, chymotrypsin, DNA polymerases, Rnase, etc. Zymogens and enzyme activation. Allosteric interactions and product inhibition; Complex kinetics and analyses, Membrane bound enzymes – Extraction, assay lipid protein interaction and effect of fludity on enzyme activity. Coenzyme; Clinical and Industrial applications of enzymes. Immobilization of enzymes and their application. Ribozymes and their applications. Enzyme engineering.

DNA transactions; replication, repair, recombination, and restriction. Plasmid biology, plasmids of gram negative bacteria, ColE1, R1, pT181, psc101. Plasmids of gram positive bacteria. P1J101. SLP and SCP plasmids of streptomyces. Plasmid vectors of various types. Biology of bacteriphage lambda. Lambda phage as natural in vivo vector, in vitro construction of a lambda vector, classes of lambda vectors. Cosmid vectors and their use. M13 vectors and their use, streptomyces phage vectors. How to choose the right type of vector. Specialized vectors- expression vectors, orf vectors, gene fusion ectors etc. Animal viruses and gene cloning. Agrobacterial plasmid biology and their use in plant genetic engineering. Vectors for yeast.

Information theory and biology; Entropy, Shannon’s formula, divergneces from equiprobability and independence, Markov chains, ergodic processes, redundancy, application to DNA and protein sequences.

Use of databases in biology: Sequence databases, structural databases, sequence Analysis- proteins and nucleic acids, structural comparisons, genome projects.

Plant genome organization, structural features of a representative plant gene, gene families in plants. Organization of chloroplast genome, nucleus – encoded and chloroplast-encoded genes for chloroplast proteins, targeting of proteins to chloroplast. Organization of mitochondrial genome, nuclear and mitochondria encoded genes for mitochondrial proteins. RNA editing in plant mitochondria, mitochondrial genome and cytoplasmic male sterility. Seed storage proteins. Maize transposable elements, organisation and function, transposable elements in transgenic plants. Regulation of gene expression in plant development. Plant hormones and phytochrome.

Symbiotic nitrogen fixation in legumes by Rhizobia – biochemistry and molecular biology, Agrobacterium and crown gall tumours. Mechanism of T_DNA transfer to plants, Ti plasmid vectors for plant transformation, Agroinfection. Classification and molecular biology of plant viruses. Molecular biology of plant stress response.

Genetic engineering in plants , selectable markers, reporter genes and promoters used in plant vectors. Direct transformation of plants by physical methods. Genetic engineering of plants for virus resistance, post resistance, herbicide tolerance, cytoplasmic male-sterility, delay of fruit ripening, resistance to fungi and bacteria. Production of antibodies , viral antigens and peptide hormones in plants, pagene silencin gin transgenic plants, DNA markers in marker-assisted selection and plant breeding. Management aspects of plant genetic Engineering. Tagging, mapping and cloning of plant genes. Molecular biology of plant pathogen interactions.

Intellectual property rights: Principles and importance.

Preparation of antigens – protocol of immunization,methods of bleeding.Preparation of serum and complement.Antigen-antibody reactions-Haemagglutination,Haemolysis,passive HA,precipitin ring test,immunodiffusion,immunoelectrophoresis,Complement fixation test,Enzyme-linked immunoabsorbent assay,Plaque forming cell assay,Isolation of immunoglobulin,Characterization,Peripheral blood mononuclear cell seperation.Lymphocyte subset identification and enumeration.Western blotting.Preparation of media,prepartion of primary culture,maintenance of secondary culture,evaluation of culture dynamics.Cell synchronization – preservation and revival of cells.Hybridoma technology and monoclonal antibody production.

Introduction to bioprocess technology parts and designs of bioreactors; production of biomass;batch and continuous fed batch fermentaion, scale up recovery of producst;Laboratory scale fermenaion of antibiotics,immobilization of cells and enzymes. Isolation of DNA from cloning vectors – plasmids and phages. Extraction of total DNA from plant and animal tissues. Isolation of restriction enzymes – demonstration of restriction systems, restriction mapping, pulsed field electrophoresis.

SEMESTER III

Core techniques in gene manipulation.Cutting and Joing DNA.Introduction of DNA into Cells.Cloning strategies, construction of genomic libraries and cDNA libraries.Probe construction, recombinant selection and screening.Analysis of expression, analysis of recombinant DNA, sequencing, mutagenesis, altered expression and engineering genes.DNA amplification using polymerase chain reaction.Key concepts.Analysis of amplified products.Applications of PCR; Ligase chain reaction.Expression systems and their applcations.E.coli.,streptomyces,Yeast,baculovirus and animal cells as cloning hosts.

Hybridoma techniques and monoclonal antibody production.Myeloma cell lines.Fusion of myeloma cells with antibody procing B-cells.Fusion methods.Selction and screeing methods for positive hybrids.Cloning methods.Production, Purification and characterization of monoclonal antibodies.Application of monoclonals in biomedical research, In clinical diagnosis and reatment.Production of human monoclonal antibodies and their applications.

T cell cloning. Mechanism of antigen recognition by T- and B- lymphocytes. Structure, function and synthesis of lymphokines. Importance of antigen presentation and MHC class II molecules in T cell cloning. Antigen specific and alloreactive T cell cloning. Use of T cell cloning in understanding the immunologically relevant antigens and T-cell epitopes. Application of T cell cloning in vaccine development.

Immunity to viruses, bacteria and parasites. Genetic control of immune response. MHC associated predisposition to diseases. Infectious diseases,Leprosy,Tuberculosis, Malaria,Filariasis,Amoebiasis,Rabis,Typhoid,Hepatitis,AIDS.Principles and strategy for developing vaccines. Newer methods of vaccine preparation. Immunodiagnosis of infectious diseases.

Intrachromosomal gene cloning in organisms other than E.coli.Degradation of toxic chemicals by pseudomonads. Protein secretion in bacteria. Biological insect control. Microbes as food and feed for animals. Microbial mining and oreleaching. Pollution and waste control. Environmental release and monitoring of genetically modified/engineered organisms.

Genetic Engineering in animals. Transformation of animal cells. Cloning vectors and expression vectors animal viral vectors and yeast vectors. Genetic engineering and applied to specific areas – regulatory proteins blood products, vaccines and hormones.

Production of usefulproteins in transgenic animals.AIDS.Oncogenes and anti-oncogenes – phasge display technogy. Signal transduction.

Baculoviruses in biocontrol and foreign gene expression. Biotechnology of aquaculture. Transgenic animals. -In vitro fertilization and embryo transfer.

Use of nucleic acid probes and antibodies in clinical diagnosis and tissue typing.Mapping of human genome. RFLP and applications. Gene therapy. Ethical issues in animal biotechnology. Management aspects of biotechnology and genetic engineering.

Paper XV-BIOPROCESS TECHNOLOGY

Isolation and screening of industrially important microbes. Improvement of the strains for increased yield and other desirable characteristics. Advantage of bioprocess over chemical process. Basic principles in bioprocess.Media formulation, strlization.Thermal death kinetics.Batch and continuous sterilization systems.Sterilization of air.Fibrous filters.Bioreactor design, parts and their functions.Types of reactors CSTR, Tower, Jet loop, Airlift, Bubble column, Packed bed.Immobilized cells.Enzyme co-immobilization.Some examples of bioprocess for the production of biomass.Primary and Secondary metabolites, extracellular enzymes, biotechnologically important intracellular products, exopolymers.

Transport phenomena in bioprocess – Mass transfer. Mass transfer coefficient for gases and liquids. Dimensionless groups. Mass transfer resistance. Rate of oxygen transfer. Determination of oxygen transfer coefficients. Theological properties of intermedium. Biological heat transfer. Heat transfer coefficients.

Bioprocess control and monitoring variables such as Temperature, agiation, pressure, pH. On line measurement. On/off control. PID control. Computers in bioprocess control systems. Downstream processing of biologicals.

DNA cloning.use of expression vectors. Southern hybridization. Preparation of radioactive and non-radioactive probes. Northern blotting and western blotting. S1 mapping. DNA sequencing. Genome analysis – bacteria, plants and animals. Site – directed mutagenesis. Polymerase chain reaction (PCR)

Micropropagation, surface – sterilization of field grown tissues, callus induction, initiation of suspension cultures, role of hormones in plant morphogenesis, regeneration of shoots and roots from callus cultures.conditioning of tissue culture plants, transfer of plants to green house.

Isolation and purification of protoplasts, viability test for protoplasts, protoplast cultures.

Induction of tumors by Agrobacterium, introduction of binary vectors into Agrobacterium by triparental mating, leaf disc/leaf transformation using Agrobacterium, GUS expression in transformed tissues, extraction of DNA from transformed plants, Southern hybridization to check plant transformation. PCR amplification of T-DNA in transformed plant tissues. Direct transformation of plant tissues by microprojectile bombardment.