North Maharashtra University, Jalgaon

Syllabus for M.Sc. Biochemistry Part II

w. e. f. 2008 - 09

Course Structure at a Glance Semester III BC-301: Plant Biotechnology BC-302: Molecular Biology BC-303: Environmental Toxicology BC-304: Lab. Course - IV BC-305: Lab. Course V (Project Work / Dissertation) Semester IV BC-401: Microbial Biotechnology BC-402: Genetic Engineering & rDNA Technology BC-403: Biochemistry of Natural Products BC-404: Applied Biochemistry & Animal Tissue Culture BC-405: Lab. Course VI
Semester I and IV will have 04 theory courses and 1 practical course each whereas semester II and III will have 3 theory courses and 2 practical courses each. Practical examination will be held annually. Each course will have a weightage of 100 marks (20 marks internal and 80 university examination) Each theory course to be completed in 50 lectures of 60 min duration each

BC-301: Plant Biotechnology

Conventional Plant Breeding: aims and objectives, methods, scope and importance. Introduction to plant biotechnology: history, concept of totipotency, heterogeneity, cytodifferentiation, preparation of explant. Tissue culture media: general introduction, composition, sterilization. Incubation systems: culture room, green house and shade house, advantages and limitations of each system. Initiation, maintenance and importance of callus culture, suspension culture and single cell culture. Plant cells as producers of secondary compounds: introduction to secondary metabolism, significance of cell differentiation, selection, downstream processing, influence of culture conditions on accumulation of secondary metabolites. Micropropagation: principle, vegetative regeneration by-shoot tip, meristem, axillary shoot production, adventitious shoot initiation, organogenesis and production of pathogen free plants. Hardening of tissue cultured plants. Somatic embryogenesis-principle, induction, importance. Artificial seeds-concept, preparation, uses and limitations. Organ Culture: principle, protocol and importance of root, stem, leaf, flower, ovary, embryo and anther culture. Protoplast culture: principle, isolation and culture of protoplast, importance of protoplast culture. Protoplast fusion-principle, methods, mechanism and importance. Somaclonal variations: concept, mechanism causing somaclonal variation, isolation and applications of somaclonal variants. Applications, scope and importance of plant cell, tissue and organ culture in agriculture, horticulture, forestry, industries.

Recommended Books; 1. Applied and fundamental aspects of plant cell, tissue and organ culture: J. Reinert and Y. P. S. Bajaj. 2. Plant cell biotechnology: R. Endress (Springer-Verlag). 3. Biotechnology in agriculture: M. S. Swaminathan. 4. Secondary metabolism in plant cell culture: P. Morrist, A. H. Scragg, A. Stafford, M. W. Fowles. 5. Agricultural Biotechnology: S. S. Purohit (Agro Botanica). 6. Plant Biotechnology: J. Hammond, P. McGravery and V. Yusibov (Eds.) (Springer-Verlag). 7. Plant cell and tissue culture for the production of food ingredients: T. J. Fu, G. Singh and W. R. Curtis (Eds.) (Kluwer Academic/Plenum Press). 8. Biotechnology in crop improvement: H. S. Chawla (International Book Distributors). 9. Plant Tissue Culture: K. K. De (New central Book Agency). 3

10. Plant cell, tissue and organ culture-fundamental methods: O. L. Gamborg and G. C. Phillips (Eds.) (Narosa Publishing House). 11. A text book of biotechnology: R. C. Dubey (S. Chand & Co. Ltd.). 12. Biotechnology: P. K. Gupta (Rastogi Publications). 13. Plant propagation-principles and practices: H. T. Hartmann and D. E. Kester (Prentice Hall India Pvt. Ltd.). 14. Practical applications of plant molecular biology: R. J. Henry (Chapman & Hall). 15. Biotechnology an introduction: S. R. Barnum (Thomson Brooks/Cole). 16. Methods in plant tissue culture: U. Kumar (Agrobios).

BC-302: Molecular Biology

DNA: Structure- current concept, triple helix DNA , melting of DNA. Replication- general principles, isolation and properties of DNA polymerases, proof reading , continuous and discontinuous synthesis. Asymmetric and dimeric nature of DNA polymerase I, II & III, exonuclease activity in eukaryotic DNA polymerases. DNA: Superhelicity, linking number, topological properties, mechanism of action of topoisomerases. DNA damage and repair: Types of DNA damage, repair pathwaysmethyl directed mismatch repair, very short patch repair, nucleotide excision repair, base excision repair, SOS repair mechanism. Transcription: General principles, basic apparatus, steps in RNA synthesis, inhibitor of transcription, control of transcription by interaction between RNA polymerase & promoter region, sigma factor, attenuation & anti termination. Maturation & processing of RNA: Methylation, cutting and trimming of rRNA, capping, polyadenylation and splicing of mRNA, catalytic RNA, group I & group II intron splicing, RNase p. Protein Biosynthesis: Steps in translation, role of various factors, various steps of translation, inhibitors of protein synthesis. Regulation of gene expression: Operon concept, catabolite repression , instability of bacterial RNA, positive & negative regulation with example, inducers & co-repressors, regulation by attenuation-his, trp, arabinose operon, autogenous regulation. Lambda phage regulation, stringent response. Protein Targeting: Synthesis of exported protein on membrane bound ribosome, signal hypothesis, SRP & its role , chaperon proteins, targeting of proteins to lysosomes, protein destruction. Eukaryotic chromosome-solenoid and nucleosomal organization, regulation of gene expression, promoters , DNA binding. Transactivator & co activators , proteins involved in transcriptional activation, regulation by intercellular & intracellular signals, translational repression, regulatory proteins.

Recommended Books; 1. Genes: B. Lewin (Oxford University Press). 2. Biochemistry: L. Stryer (W.H. Freeman & co.). 3. Lehninger principles of biochemistry: D. L. Nelson & M. M. Cox (Macmillan Worth Publishers). 4. Molecular cell biology: J. Darnell, H. Lodish & D. Baltimore (Scientific American Books). 5. Fundamentals of biochemistry: D. Voet, J. Voet & C. Prati (John Wiley & Sons). 6. Molecular biology of genes: J. Watson, N. H. Hopkin, J. W. Roberts, J. P. Stertz & A. M. Weiner (Freeman Co.) 5

7. The biochemistry of the nucleic acids: R. L. P. Adams, J. T. Knowler & D. P. Leader (Chapman & Hall). 8. Biotechnology an introduction: S. R. Barnum (Thomson Brooks/Cole). 9. Cell and molecular biology: E. D. P. De Robertis, E. M. F. De Robertis, Jr. (Lea & Febiger international edition, Info Med). 10. Principles of genetics: E. J. Gardner, M. J. Simmons, D. P. Snustad (John Wiley & sons Inc.). 11. A history of molecular biology: M. Morange (Oxford University Press). 12. Cell and molecular biology: P. Sheeler, D. E. Blanchi (John Wiley & sons Inc.). 13. Molecular biology and microbes: S. Srivastava and V. Singhal (Anmol publications Pvt. Ltd.). 14. Concepts in molecular biology: S. C. Rastogi, V. N. Sharma & A. Tandon (Wiley Eastern Ltd.). 15. Molecular biology: P. K. Gupta (Rastogi Publications).

BC-303: Environmental Toxicology

Principles of toxicology: Basis for general classification and nature, definition and purpose of toxicology, dose response relationship, synergism and antagonism. Determination of ED50, LD50 and chronic exposures. Factors influencing toxicity. Principles and procedures of testing for acute toxic effects: Regulatory guidelines, mammalian system affected and the clinical signs of systemic toxicity. Factors affecting acute toxicity studies. Xenobiotic metabolism: Absorption and distribution, phase I reactions - oxidation, reduction, hydrolysis, hydration. Phase II reactions - conjugation, methylation, glutathione and amino acid conjugation. Detoxification. Toxicity testing: Test protocol, genetic toxicity testing and mutagenesis assays, in vitro test systems, bacterial mutation tests, preservation test, Ames test, in vivo mammalian mutation tests, host mediated assay and dominant lethal test. Use of Drosophila in toxicity testing. DNA repair assays, chromosome damage test, toxicological evaluation of recombinant DNA derived proteins. Biological effect of ionizing radiation and toxic effects of animals and plants toxins: Radiation carcinogenesis, acute radiation syndrome, properties and effects of animal toxins snake and scorpion, plants toxins ricin and mycotoxins (aflatoxin B1 & G1). Pesticide toxicity: Insecticides - organochlorines, anticholinesterases, organophosphates, carbamate. Fungicides and herbicides. Environmental consequences of pesticide toxicity. Biopesticides general introduction, scope and importance. Metal toxicity: Toxicology of Arsenic, Mercury, Lead and Cadmium. Environmental factors affecting metal toxicity. Effects of light, temperature and pH. Toxicology and detoxification of pollutant: Common air pollutants and their sources. Air pollution and Ozone, air pollution due to chlorofluorocarbons (CFCS) and asbestos. Control of air pollution. pollutants sources, adverse effects and control measures. Detoxification Solid waste management: toxic effects and treatments, methods, technologies for management of hospital waste incineration, autoclaving, mechanical/chemical, microwave, plasma torch, detoxification, advanced wet oxidation and thermal dry heat.

Recommended Books; 1. Casarett and Doulls toxicology: C. D. Klaassen, M. O. Amdur, J. Doull (Macmillan Publishing Co.). 2. Hayes principles and method of toxicology: A. Wallace and Hages (Eds.) (Raven Press). 3. Hamilton and Hardys industrial toxicology: R. D. Harbison (Tampa). 4. Basic toxicology fundamental target organs and risk assessment: F. C. Lu. 5. Environmental biology: K. C. Agrawal (Agor Botanica). 7

6. Toxicology: A. Sood (Sarup & sons). 7. Introduction to environmental toxicology impact of chemicals upon ecological systems: W. G. Landis and Ming-Ho Yu (Lewis Publishers Inc.). 8. Hamilton and Hardys Industrial toxicology: R. D. Harbison (Mosby). 9. CRC handbook of toxicology: M. J. Derelanko, M. A. Hollinger (CRC Boca Raton). 10. Environmental toxicology human exposure and their health effects: M. Lipmann (John Wiley & sons Inc.). 11. Principles and methods of toxicology: A. W. Hayes (Raven Press). 12. Toxic and hazardous waste disposal: R. B. Pojasek (Ann Arborscience Publisher). 13. Toxics A to Z a guide to everyday pollution hazards: J. Harte, C. Holdren, R. Schneider and C. Shinley (Bishen Singh Mahendrapal Singh Publishers). 14. Environmental toxicology: M. Satake, Y. Mido, H. Yasuhisa, S. Taguchi, M. S. Sethi, S. A. Iqbal (Discovery Publishing House). 15. Fundamental toxicology for chemists: J. H. Duffus and H. G. J. Worth (Eds.) (The Royal Society of Chemistry). 16. Waste Management: R. K. Sinha and A. K. Sinha (INA Shree Publishers).

BC-304: Laboratory Course - IV

1. Plant tissue culture laboratory organization. 2. Comparative efficacy of chemical sterilents for sterilization of plant material. 3. Induction and characterization of callus. 4. Isolation of secondary metabolite from the callus tissue of medicinally important plant. 5. Regeneration (caulogenesis and rhizogenesis) of plant from callus. 6. Micropropagation through shoot tip/meristem culture. 7. Regeneration of high yielding banana plantlet from rhizome. 8. Somatic embryogenesis from suitable explant. 9. Preparation and evaluation of microbial biopesticide. 10. Preparation and evaluation of botanical biopesticide. 11. Isolation and determination of bioefficacy of phosphate solubilising microorganisms. 12. Isolation and determination of bioefficacy of nitrogen fixing microorganisms. 13. Determination of ED50 and LD50 of a commonly used pesticide/s. 14. Isolation and estimation of alkaloid/steroid/flavanoid from suitable source (D). 15. Seperation of plant secondary metabolites by HPTLC (D). (Any ten experiments to be performed.) Recommended Books; 1. 2. 3. 4. 5. 6. An introduction to plant tissue culture: Kalyan Kumar De (New Central Book Depot). Plant cell and tissue culture- a laboratory manual: J. Reinert and M. M. Yeon. Plant tissue culture- techniques and experiments: R. Smith. Methods in plant molecular biology: M. A. Schuler and R. E. Zielinski. A laboratory manual of plant biotechnology: S. S. Purohit (AgroBotanica). Methods in plant tissue culture: U. Kumar (Agrobios).

BC-305: Laboratory Course V (Project Work / Dissertation)

BC-401: Microbial Biotechnology

Introduction to fermentation technology: The range of fermentation processes, microbial biomass, microbial enzymes, recombinant products, transformation products. Chronological development of fermentations industry. Screening of organism: Primary , secondary screening, modern concept for screening. Nisbet guideline for overproduction media. Preservation and improvement of industrially important organisms: Criteria for choice organism, selection of autotrophic mutants, morphological mutant, analogue, precursor, resistant mutant, recombinations. Preservation methods, low temperatures, dehydration, quality control of preserved cultures, culture collection centers. Medium for industrial fermentations: Criterias for designing fermentation medium formulation, cellular yield coefficient, media component , carbon sources, nitrogen source, minerals, antifoams, precursors, inducers and inhibitors. Media optimization by using Placklett Burmen design. Inoculum development for industrial fermentation, criteria to inoculate organism, criteria for transfer of inoculum. Development of inoculums for yeast, fungi, actinomycetes and bacteria. Sterilization of industrial fermentation medium: Methods of sterilization - steam, radiation, filtration, gaseous. Batch sterilization, designing of batch and filtration. Continuous sterilization. Calculation of Del factor, Richard rapid method for designing of sterilization cycle. Industrially significant fermentations: Antibiotics - penicillin, tetracycline. Vitamins riboflavin biotin. Organic acids - citric acid, gluconic acid. Alcohols - ethanol, butanol. Solid state fermentation: advantages, disadvantages, comparison with submerged fermentation, bioreactors used for solid state fermentations. Dual fermentation examples, need and significance. Applications of industrially important enzymes: lipases, amylases, pectinases, hylaluraunidase. Introduction to some industrially important products - serretia, peptidases, erythropoietin, rifamycins, nisin, lovostatins. Downstream processing: Cell separation, disintegration, extraction liquid -liquid, two phase liquid. Precipitation, purification, crystallization, spray drying, drum dryers, packaging. Specific examples of fermentation product recovery.

Recommended Books; 1. Bitechnology of filamentous fungi: D. B. Franklinstein and C. Ball (BitterworthHeinemann). 2. Biotechnology of antibiotics: Vandamme Drugs and pharmaceuticals, Vol. 22 (Ed.) (Marcel Dekker Inc.). 10

3. Principle of fermentation technology: P. F. Stransberry, A. Whitekar & S. J. Hall ( Aditya Books). 4. Process biotechnology fundamentals: Mukhopadhay (Viva Books Pvt. Ltd.). 5. Microbial technology - fermentation technology, Vol. I & II: A. H. J. Peppler & D. Perlmann (Academic Press). 6. Solid substrate cultivation: H. W. Doelle, D. A. Mitchell & C. E. (Elsevier Applied Science). 7. Industrial microbiology: L. E. Casida JR (New Age International Publishers Ltd.). 8. Industrial microbiology: S. C. Prescott & C. G. Dunn (Agrobios). 9. Biotechnology an introduction: S. R. Barnum (Thomson Brooks / Cole). 10. Basic biotechnology: C. Ratledge and B. Kristiansen (Cambridge University Press).

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BC-402: Genetic Engineering and rDNA Technology

Genetic engineering: Concept. Restriction enzymes nomenclature, properties, types, mode of action. Restriction modification system. Cloning vectors: Plasmids, bacteriophages, cosmids, phagemids. chromosomes. Binary and shuttle vectors, expression Yeast cloning vectors, cloning in yeast. Artifical vectors.

Cloning: Steps involved, strategies for each step with details of cDNA synthesis of generation of DNA, joining cDNA to vector DNA by homopolymer tailing , transformation with rDNA, transfection with r-phage DNA, screening by immunochemical and nucleic acid hybridization method. DNA library: cDNA and genomic DNA library. Mapping: S1 nuclease mapping, restriction mapping and their applications. Mutagenesis: Site directed mutagenesis and its applications. Polymerase chain reaction, RFLP, RAPD, AFLP and their applications. Southern blotting. DNA sequencing enzymatic, chemical and automated. Transformation in plants: feature of Ti and Ri plasmid, basis of tumour formation, mechanism of DNA transfer, role of virulence geness, use of reporter genes. Viral vectors and their applications, particle bombardment, electroporation, microinjection. Application of plant transformation for productivity, performance, herbicide resistance, long shelf life of fruits and vegetables. An introduction to markers, quantitative trait loci (QTL) mapping. Marker assisted selection for crop improvement. DNA chip technology and microarrays: Types and production of chips. Applications of microarrays on DNA chips. RNAi. Gene silencing and antisense technology. Gene therapy - Vectors for gene therapy, gene editing and gene replacement. Scope and applications of genetic engineering in medicine, agriculture and industry. Biosafety: Guideline for safety in biotechnology, good laboratory practice (GLP), good large scale industrial practice (GLSP), biosafety level for infectious agents. Regulations environment, environmental pollution and hazardous substance.

Recommended Books; 1. Recombinant DNA: J. D. Watson, M. Gilman, J. Witkowski and M. Zoller (Scientific American Books distriduted by W. H. Freeman and co.). 2. Principles of gene manipulation an introduction to genetic engineering: R. W. Old and S. B. Primrose (Blackwell Scientific Publications). 3. Genes: B. Lewin (Oxford University Press). 4. Biochemistry: L. Stryer (W.H. Freeman & co.). 5. Lehninger principles of biochemistry: D. L. Nelson & M. M. Cox (Macmillan Worth Publishers). 6. Cell and molecular biology: P. Sheeter and D. E. Blanchi (John Wiley & sons Inc.) 7. Methods in molecular biology, Vol. 92 PCR in bioanalysis: S. J. Metzer (Humamna press). 8. Gene function E. coli and its heritable elements: R. C. Glass (Croom Helm). 12

9. Principles of genetics: E. J. Gardner, M. J. Simmons, D. P. Snustad (John Wiley & sons Inc.). 10. Genetic engineering: R. Williamson (Academic press Inc.). 11. Molecular cloning a laboratory manual : J. Sambrook, E. F. Frithsch and T. Maniatis (Cold Spring Harbor Laboratory Press). 12. Molecular Biotechnology: S. B. Primrose (Blackwell Scientific Publication). 13. Methods in enzymology Vol. 152; guide to molecular cloning techniques (Academic Press). 14. DNA cloning a practical approach: D. M. Glover and B. D> Hames (IRL Press). 15. Gene therapy principles and applications: T. Blankenstein (Birkhauser Verlag). 16. Genetic engineering: P. K. Gupta (Rastogi publications). 17. Biotechnology and genomics: P. K. Gupta (Rastogi publications). 18. Biotechnology: R. C. Dubey (S. Chand). 19. The Indian environment protection act, 1986 (Govt. of India). 20. Rules for manufacture, use / import / export / and storage of hazardous microorganisms or cells act, 1989 (Govt. of India). 21. Food safety and standards act. 2006 (Govt. of India).

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BC-403: Biochemistry of Natural Products

General aspect of sources of natural medicinal plant products: Introduction to primary and secondary metabolites, types of secondary metabolites, production under stress, isolation of active constituent from plant material. Alkaloids: Definition, general properties, classification based on nitrogen heterocyclic ring, types - phenylalkylamines, pyridine alkaloids, tropane alkaloids, quinolizidine and pyrrolizidine alkaloids, isoquinoline alkaloids, quinoline, monoterpene, indole alkaloids, purine alkaloids, ruta alkaloids, medicinal importance of each type. Role of alkaloids in plants. Saponins and Steroids: Definition, general properties, medicinal importance of saponins. Important saponins of plant origin - diosgenin, hecogenin, glycyrrhizin, aescin and ginseng. Steroids: Definition, general properties, classification, introduction and medicinal importance of - cardiac glycosides from Digitalis, Strophanthus, Urginea, steroids from Withania somnifera, Holarrhena and Solanum. Terpenoids: Definition, general properties, classification, introduction and medicinal importance of terpenoids. General account and medicinal importance of myrcene, ocimene, citronellol, menthol and camphor. Tannins, lignin and pectin: Definition, general properties and classification. Terpenes: Definition, general properties and classification. Plant pigments: Occurrence, classification, introduction and applications of carotenoids, xanthophylls, anthocyanins, flavones, flavonols. Acetate pathway and Shikimic acid pathway. Pyrethroids and rotennones of plant origin: importance. Definition, general properties and

Intellectual property rights: Patents, trade secrets, copyrights, trademarks and geographical indications. IPR and plant genetic resources. IPR for plant breeding Sui Generis system, plant variety protection and UPOV, plant variety protection in India and USA. Plant breeders rights and farmers rights.

Recommended Books; 1. Biotechnology - secondary metabolites: K. G. Ramawat and J. M. Merillon (Eds.) (Oxford & IBH publishing Co. Pvt. Ltd.). 2. Natural products - chemistry and biological significance: J. Mann, R. S. Davidson, J. B. Hobbs, D. V. Banthrope and J. B. Harbonne (Longman). 3. New trends in natural product chemistry: Ata-ur-Rahman and M. L. Choudhary (Harwood Academic Publishers). 4. Pharmacognosy: G. E. Trease and W. C. Evans (ELBS) 5. Text book of pharmacognosy: T. E. Wallis (CBS Publishers) 14

6. Organic chemistry of natural products: Gurdeep Chatwal (Himalaya publishing House). 7. Indian medicinal plants: Kirthikar and Basu 8. Materia medica: Henry G. Greenish (Scientific Publishers) 9. Agrochemicals from natural products: C. R. A. Godfrey(Ed.) (Marcel Dekker, Inc.) 10. The Total synthesis of natural products: John ApSimon (Ed.) (John Wiley & Sons, Inc.). 11. Introduction to flavonoids: B. A. Bohn (Harwood Academic Publishers). 12. Insecticides of natural origin: Sukh Dev (Harwood Academic Publishers). 13. Chemistry and biology of herbal medicine: V. P. Agrawal and V. P. Khamboj (Eds.) (Society of Biosciences). 14. Foyes Principles of medicinal chemistry: T. L. Lemke, D. A. Williams, V. F. Roche and S. W. Zito (Wolters Kluwer / Lippincott Williams & Wilkins). 15. Biopharmaceueticals - biochemistry & biotechnology: G. Walsh (Wiley).

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BC-404: Applied Biochemistry and Animal Tissue Culture

Pharmaceuticals Biochemistry: Introduction, theories of drug action, drug receptors, isolation of drug receptor, rate theory, induced fit theory, theory of macromolecular perturbation. Mechanism of drug action, action of enzymes, drug acting as suppressor of gene function. Drugs acting on biological membranes, non specific action of drugs. Metabolism of drug: Assay of drugs, phases of metabolic processes of drugs, different type of drug receptors, drug absorption and factors affecting drug metabolism. Different types of drugs: Anesthetics, tranquillizer, diuretics, anti-malerial, anti inflammatory and cardiovascular agents. Anti-AIDS drug development, anti nutrients naturally occurring food borne toxicants, protease inhibitors, hepatic toxins, allergens, oxalate toxins from mushroom. Nanobiotechnology: Concept, Scope and applications - biological applications of carbon nano tubes, biosynthesis of metals, non metal and magnetic nano particles, bioremediation and nano medicines. Clinical biochemistry: Introduction to stems cells, disorders of carbohydrate metabolism, disorders of aminoacids metabolism. Clinical significance of SGOT, SGPT, CK and LDH. Inborn errors of metabolism. Animal cell culture: Introduction, application and limitation. Equipments used in tissue culture, glassware and plasticware used in animal tissue culture. Aseptic techniques in animal tissue culture. Tissue culture media. Types of tissue culture - primary cultures, epidermal, corneal, cervical, tumors cell, cerebellar, chondrocytes, osteoblast, melanocytes and bone marrow. Culture techniques: Transformation, differentiations, dedifferentiation. Culture cell lines, initiation and evolution of cell lines, maintenance, phases of cycle, growth curves. Protocols for subcultures - monolayer and suspension cultures. Types of contaminations. Protocols for routine characterizations of cell lines. Specialized techniques in animal tissue culture, hybridoma cultures, stems cell culture.

Recommended Books; 1. 2. 3. 4. 5. Culture of animal cell: R. I. Freshney (Wiley - Liss). Genes: B. Lewin (Pearson Educational International). Animal cell culture Practical approach: R. W. Jhon (Masters Oxford). Biotechnology: U. Satyanarayana (Books & Allied Pvt. Ltd.). Methods in cell Biology - Vol. 57 - animal cell culture methods: J. P. Mathon and D. Barnes (Eds.) (Academic Press). 16

6. Synthetic drugs: M.S. Yadav. 7. Synthetic organic chemistry: Gurudeep Chatwal (Himalaya Publishing House). 8. Foyes Principles of medicinal chemistry: T. L. Lemke, D. A. Williams, V. F. Roche and S. W. Zito (Wolters Kluwer / Lippincott Williams & Wilkins). 9. Biopharmaceueticals - biochemistry & biotechnology: G. Walsh (Wiley). 10. Biotechnology and Pharmacy: J. M. Pezzuta, M. Jhonson and H. R. Manasse.

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BC-405: Laboratory Course - VI

1. Isolation of chromosomal / genomic DNA from Bacteria / Yeast / plant / animal cell / tissue. 2. Isolation and characterization of plasmid DNA. 3. Estimation of DNA by DPA method. 4. Estimation of RNA by orcinol. 5. Agarose gel electrophoresis of DNA. 6. Induction of lac operon in E. coli. 7. Determination of Tm of DNA. 8. Determination of G+C content of DNA. 9. Restriction digestion of DNA and analysis by agarose gel electrophoresis. 10. Amplification of DNA by polymerase chain reaction (PCR). 11. Isolation of Bacteriophage by plaque assay. 12. Seperation of peptides by PAGE. 13. Preparation of competent bacterial cells and their transformation by using plasmid. 14. Agrobacterium mediated transformation of plant material (root / tumour induction). 15. Demonstration of Southern blotting technique. (Any ten experiments to be performed) Recommended Books; 1. Molecular cloning a laboratory manual Vol. I, II & III: Sambrook and Russel (Cold Springer Harbour Laboratory Press). 2. Practical biochemistry - principles and techniques: K. Wilson and J. Walker (Cambridge Press). 3. An introduction to practical biochemistry: D. T. Plummmer (Tata Mc Hill). 4. Methods in agriculture al Biochemistry: S. Sadashivam and A. Manikam (New Age Publication). 5. Laborotary manual in general Microbiology: H. J. Binson (Wm C Brown Publishers). 6. Laboratory manual in biochemistry: J. Jayraman (New Age Publication Ltd.) 7. Lab exercises in microbiology: J. P. Harly and L. M. Prescott (WBC / Mac Hill).

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