PAPER VI Bacterial Genetics and Molecular Biology
Unit 1 :[I] Introduction to bacterial genetics.
A. Principles of inheritance: relevance of Mendelian laws.
B. Nature of genetic material, gene structure and function, arrangement and linkage, gene complementation, cistron, concept of intron
and exon. one gene one poly peptide theory, organisation of bacterial chromosome,microorganisms as a genetic tools.
[II] Replication of DNA.
A. Semi-conservative mode of chromosome replication, work of Messelson and Stahl.
B. Molecular mechanism of chromosome replication, origin of replication,mode of formation of replication fork and its growth, post
replicativemodification of DNA.
C. Models of chromosome replication - Cairn's model and Rolling circle model.
Unit 2 : Gene expression and its regulation.
A. Concept of central dogma.
B. Gene transcription as the first step of gene expression, molecular
mechanism of transcription, role of RNA polymerase, initiation,
elongation and termination of RNA synthesis, post transcriptionalmodifications in bacteria.
C. Nature of Genetic Code: characteristics of genetic code, triplet nature,degeneracy, and wobble hypothesis Deciphering genetic
code, work ofNirenburg. Holley and Khorana.
D. Gene translation:
1. Ribosomes: their general nature, structure and role in protein synthesis
2. Type of RNAs involved in protein synthesis, structure and function of tRNA
3. Initiation, elongation. & termination of protein synthesis; post translational processing.
4. Protein localisation: export of protein, role of signal peptides.
E. Regulation of gene expression:
1. Operon model.
2. Regulation of inducible and constitutive genes.
3. Positive Sc negative control, catabolic repression, variation in. sigma factor & promoter in gene regulation.
4. Introduction to other patterns of gene regulation.
Unit 3 : Mutation and DNA repair.
A. Nature of mutation, spontaneous and inducible mutation, mutation rate.
Mutagens; phenomic and phenotypic lag. -
Phenotypic classes of bacterial mutants. Methods of their isolation.
B. Molecular basis of mutagenesis.
Mode of action of some mutagens, base analogues, nitrous acid, alkylating agents, UV rays, Mu phage as a mutagen. Ame's
test.
Reversion of mutation, true reversion and suppression, types of suppressor mutations.
C. DNA repair: direct and indirect repair mechanisms, post replicative and recombinational repair.
Unit 4 : Genetic recombination and gene transfer in bacteria.
A. Introduction to genetic recombination and its biological significance,types of recombination and their molecular mechanisms:
generalised, sitespecific and illegitimate recombination, recombination frequency and itssignificance.
B. Modes of genetic transfer in bacteria: merodiploidic nature of bacterial zygote.
Transformation: transformation principle, competence factor, mechanism of DNA uptake, transfection.
Transduction: phages involved,, types-restricted, generalised and abortive transduction
Conjugation: role of sex factor, types of crosses involved, F+ and Hfr cells. Mechanism of chromosomal transfer, interrupted
mating and its applications, zygotic induction, sexduction.
Plasmids and transposable elements as tools of gene transfer.
Unit 5 : rDNA technology and genetic engineering.
A. Introduction to genetic engineering, gene cloning and its ethical consideration.
B. Outlines of rDNA technology and its application.
C. Tools of genetic engineering/rDNA technology.
Enzymes: restriction endonucleases, RNA polymerase. DNA ligase, alkaline phosphatase, reverse transcriptase.
Cloning vectors - plasmids, bacteriophages, cosmids, Ti plasmid, yeast episomal plasmid (YEP).
Genetic probe and PCR.
D. Basic techniques of gene cloning in prokaryotes: brief outline.
E. Site directed mutagenesis.
TEXT BOOKS. 3
1. Stanier R.Y.. ingraham. J.L. Wheelis. M.L Painter. PR., (1986) General . Microbiology. (5th ed.) Mac Millan Press Ltd. London.
2. Atlas, R.M (1997) Principles of Microbiology. (2nd edn). Wm. C. Brown Publishers. 4 Dubuque.
3. PrescotL L.M.; Harley, J.P., Klein. DA.. (2002) Microbiology (5th ed), McGraw Hill, International ed.
4. Trevan, M.D., Boffey, S.. Goulding, K.H., Stanbury, P., ,1988) Biotechnology: The' Biological Principles. Tata Mcgraw-Hill, New Delhi.
5. Madigan, M.T.. Martinko, J.M., and Parker. J. (2003) Brock Biology of Microorganisms. ClOth edV Prentice-Hall International NJ USA
PAPER VII BACTERIAL METABOLISM
Unit 1 : Energy, enzyme and regulation.
A. Enzyme:
Enzyme kinetics - Michalis & Menten equation, Lineweaver-Burk plot, Kra value, Vmi!(, importance of Km and VIM:c in understanding
en7.yme reaction.
Enzyme regulation: the nature and significance of metabolic regulations, metabolic channelling.
Allostarit regulation, allosteric site and its role.
Feed back inhibition of metabolic pathways, patterns of regulation of branched chain pathway.
Other methods of control of enzyme activity - energy linked control, precursor activation, zymogen activation.
B. Energy:
Laws of Thermodynamics.
Free energy and reactions, E'O value and its importance.
Role of ATP in metabolism.
Oxidation-reduction reactions and electron carriers.
Unit 2 : Energy release and conservation.
A. Heterotrophic mode.
Breakdown of glucose under aerobic and anaerobic conditions.
TCA cycle and glyoxal bypass.
Aerobic hydrocarbon degradation: β-keto adipate pathway.
Modes of ATP formations.
Electron transport and oxidative phosphorylation, mechanism of generation of proton motive force.
Anaerobic respiration.
Catabolism of carbohydrate and intracellular reserve polymers.
Lipid catabolism: β-oxidation pathway.
Protein and amino acids catabolism.
B. Chemoautotrophic mode.
Oxidation of inorganic molecules: hydrogen, iron, sulphur and nitrogen.
Role of reverse electron transport in generation of reducing power.
Photosynthesis: light reactions, comparative account of energy release in plant and bacterial photosynthesis
Unit 3 : The use of energy in biosynthesis.
Principles governing biosynthesis.
Photosynthetic fixation of carbon dioxide.
Gluconeogenesis.
Assimilation of inorganic sulphur and nitrogen.
Anaplerotic reactions.
Lipid synthesis- synthesis of fatty acids and phospholipids.
Peptidoglycan synthesis.
Unit 4 : Separation and analysis of metabolites and metabolic pathways.
A. Principles and applications of:
Separation methods: centrifugation, chromatographic, electrophoretic.
Analytical methods: spectroscopic.
B. Methods of studying biosynthesis:.
Isotopes and pulse labelling.
Biochemical mutants.
Metabolic inhibitors.
Unit 5 : Biostatistics and bioinformatics.
A. Biostatistics.
Introduction of biostatistics, data and sampling.
Graphical representation of data.
Measures of central value: mean, median and mode.
Measure of variability of data: standard deviation.
B. Bioinformatics.
Introduction
Importance and applications of bioinformatics.
Computer: basics, basics of hardware and software.
Uses of computer in biology and bioinformatics.
TEXTBOOKS.
1. R.Y. Stanier. Adelbera. E.A.. Ingraham, J.: (1986) The microbial world. (5th ed), Prentice-Hall. N.J.
2. Atlas. R.M. (1997) Principles of Microbiology. (2nd edn). Wm.C. Brown Publishers. Dubuque
3. Prescott, L.M., Harley. J.P., Klein, D.A., (2002) Microbiology (5th ed), McGraw Hill, International ed.
4. Agricultural Statistics Techniques and Procedure, Mandal and Nambiar, 2002
5. Clarke, G.M. (1994) Statistics and Experimental Design: An Introduction for Biologists and Biochemists. (3rd ed) Edward Arnold, London.
6. Introduction to Microbioogy, J. L. Ingraham and C. A. Ingraham, 2000.
PAPER VIII IMMUNOLOGY AND CLINICAL MICROBIOLOGY
Unit 1 : Immunity and immune response.
A. Immunity: introduction; types of immunity - innate and acquired, active and passive, natural and artificial, herd immunity.
B. Immune response.
Antigen and immunogenicity: definition, character of immunogens, antigenic determinants, adjuvants, types of antigens, bacterial
antigens.
Immunoglobulins: general characters, basic structure of immunoglobulins, classes of immunoglobulins and their physicochemica
and biological characteristics.
Monoclonal antibodies and their applications.
Immune response and immune system: nature of immune response, primary and secondary immune response, factors affecting
immune response.
Immune systemperipheral and central lymphoid system. Lymphoid organs. Cells involved in immune response. Clonal selection
theory, basis of antibody diversity.
Unit 2 : Dysfunctional immunity.
A. Introduction, types, autoimmune disorders.
B. Immunodeficiency: congenital and acquired.
C. Hypersensitivity introduction and types of hypersensitivity.
D. Transplantation immunity: MHC antigens. Host vs graft and graft vs host reaction, immuno suppression.
E. Tumour immunity.
Unit 3 : Diagnostic immunology and prophylactic immunisation.
A. Diagnostic immunology
Antigen antibody reactions:, types, mechanism, zone phenomenon
In vivo antigen-antibody reactions, immune complex formation. Complement fixation: classical and alternate pathways.
Neutralisation, virus neutralisation, Opsonization.
In vitro antigen antibody reactions: precipitation, agglutination, complement fixation. ELISA, Rl A, RAST, Immunoflourescence.
Western Blot.
Measurement of CMI IRMIF and MLR.
Skin tests.
B. Prophylactic immunisation: introduction, characteristics and types of vaccines, schedule of VHCcination. hazards of vaccination.
Unit 4 : Host parasite relationship and epidemiology.
A. Normal flora of body: role, origin and establishment: normal flora of different systems; germ free animals and gnotobiosis
B. Host Parasite interactions: dynamicity of host parasite relationship; infection, factors affecting infection, infective process, types of
infection,nosocomial infections.
C. Virulence of pathogenic organisms: Microbial factors: invasiveness and toxigencity.
D. Host defenses: non-specific host defenses, general barriers-physical, chemical and biological: specific host defenses.
E. Epidemiology and types of infection
Unit 5 : Clinical microbiology.
A. General symptoms, transmission and control of bacterial infection of: skin, eye, respiratory tract, cardiovascular system, nervous
system, gastrointestinal tract, urinogenitial tract
B. Clinical microbiology:
Specimens: collection, handling, transport.
Identification of microorganisms from specimens -microscopic, culturalgrowth & biochemical characteristics; phage typing,
susceptibility testing.
Metabolic products (pathological changes in hood, body fluids & tissues as indicators).
Serological/immunological techniques.
Rapid identification techniques, molecular methods.
C. Haematology:
Blood: its components, structure and function of blood cells, blood cell maturation
Blood coagulation.
Human blood group system.
Principles of blood banking and safety in blood transfusion.
TEXT BOOKS
1. Atlas, R.M. (1997) Principles of Microbiology. (2nd ed.). Win. C Brown Publishers. Dubuque.
2. Prescott, L.M., Harley, J.P., Klein. DA., (2002) Microbiology (5th edY McGraw Hiil. International ed.
3. Tortora, G.J., Funke, B.R., Case, C.L. (2001) Microbiology: An Introduction. (7th ed). Benjamin Cummings N.Y.
4. Baker, F., Silverton, R.R. Pallister C..T. (1998) Baker and Silverton's Introduction to Medical Laboratory Technology. (7th edn.).
Butterworths. Heinemann. Oxford. UK
PAPER IX : ENVIRONMENTAL MICROBIOLOGY.
Unit-I: Ecological aspects of Biodegradation. control and soil, water, and water management technology.
Control of biodegradation.
Management of Aquiculture soils
Fouling biofilms.
Treatment of solid-waste
Landfields - composting
Treatment of liquid waste.
1. Biological Oxygen Demand
2. Fixed films sewage Treatment system
3. Suspended cell: sewage Treatment system
4. Tertiary treatments - Disinfections treatment and safety of water supplies,
5. Water quality testing.
Unit-II Microbial interreaction with Xenobiotic and inorganic pollutants.
Persistance of Biomagnification of Xenobiotic molecules.
Recalcitrant Helocarbons.
Recalcitrant Nitroaeromatic compounds
Polychlorinated biophenyl and Dioxins,
Synthetic polymers,
Alkyal Benzyl sulfonates
Petroleum hydrocarbons
Pesticides
Heavy metals and Radionuclides.
Unit - III (A) Microbial adaptation
Abiotic limitation to microbial growth
Liebig law of the minimum.
Shelfeild's law of minimum.
Starvation strategies
Environmental determinants
Effect of various factors on microbial activity.
(B) Microbial adaptation to environment and Biodegradability testing.
Biodegradability and ecological side effect testing
Testing for biodegradability and Biomagnifications
Testing for effect on microorganism
Approaches to bioremediation
Bioremediation of Marine oil pollutant
Bioremediation of spills Exxon Valdez Alaban oil pollutant.
Bioremediation of air pollutants
Unit - IV Microorganism in mineral & Energy Recovery and Fuel and Biomass production
Recovery of metals
Microbial assimilation of metal
Bioleaching of copper and Gold
Uranium Bioleaching
Phosphate recovery
Recovery of petroleum
Production of fuels Ethanol, Methanol, Hydrocarbons
Other then Methane and hydrogen
Production of microbial biomass
Single Cell Protein production
Mushroom production
Unit - V (A) Microbial diseases of plant
Viral diseases of plants, TMV, Potato spindle
Bacterial diseases of plants, Ervinia Agrobaterium.
Fungal diseases of plants: Rust, Blight, Smut of cereals
Mycorrhiza.
Ecto Mycorrhiza and endo Mycorrhiza
Application
(B) Microbial control
Microbial control of insect peats
Microbial control of weeds and cyanobacterial bloom.
Bacterial control of Dutch Elm dieaso.
Genetic engineering in biological control.
Forest protection.
Bacillus thuringiensis in pesticides
Other application.
PAPER X FERMENTATION BIOTECHNOLOGY
Unit I : Fermentation technology - I
A. Introduction to fermentation processes, concept of fermentation, range of fermentation processes, component parts.
B. Media for fermentation: raw materials used, formulation of media, criteria for selection on raw materials, media ingredients.
C. Sterilization:
Need for asepsis protected fermentation.
Media sterilization-use of high pressure steam, i) value and its significance factors affecting: batch and continuous sterilization
Air sterilization -theory of sterilizations, methods, types of fillers
Sterilization of fermenter introduction to filter sterilization of media and exhaust air.
Principles and methods of development of seed culture: general principles involved in preparation of bacterial and fungal inoculum
D. Immobilisation of cells and enzvmes.
Unit 2 : Fermentation technology - II
A. Bioreactors:
Basic (functions, aseptic operation and containment.
Aeration and agitation components: needs for aeration and agitation, (actors affecting its efficiency. KLa and its significance)
Achievement and maintenance of asepsis
B. Introduction to scale up
C. Down stream processing:
Introduction.
Removal of microbial cells and suspended solids.
Introduction to principles of cell disruption methods: physical, chemical and enzymatic methods.
Concentration by solubilization, solvent extraction and precipitation.
Purification by crystallisation, chromatographic methods and ultrafilteration
Drying.
Quality assurance bioassay
D. Introduction to fermentation economics.
Unit 3: Isolation, preservation & improvement of industrially important microorganisms.
A. Criteria for selection of industrially important organisms.
B. Screening methods: primary and secondary screening.
C. Principles involved in isolation methods.
D. improvement of industrially important microorganisms:
Need for strain improvement.
Strategies for strain improvement: selection, adaptation, mutation and rDNA techniques.
Unit 4 : Microbial processes: I
A. Microorganisms involved, fermentation processes and product recovery of:
penicillin, ethanol, citric acid, vitamin B12. lysine, xanthan. B Biomass production: production of food and feed yeast.
Unit 5 : Microbial processes: II
A. Microorganisms involved, processes and products:
Microbial production of enzymes - amylase and protease;
Applications of enzymes - therapeutic, analytical, manipulative, and industrial.
Bioextraction of metals.
Microbiallly enhanced oil recovery (MEOR)
Microbial insecticides.
Biofertilizers.
TEXT BOOKS.
1. Stanbury. P.lv, Whitaker. A., Hall. S. J.(199.V) Principles of Fermentation Technology. (2nd edni Butterworth Heinemann Ltd. Oxford, U.K.
2. Trevan. M.D., Boffey. S.. Goulding. K.H., Stanbury. P., (1988) Biotechnology: I he Biological Principles Tata Mcgraw-Hill, New Delhi.
3. Atlas. R.M. i'1997'i Principles of Microbiology. (2nd edn). Win. C. Brown Publishers Duhuque.
4. Cruger. VV. and Cruger. A (1990) Biotechnology: A Textbook oj Industrial
Microbiology. ('2nd ed), ed. T.D.Brock Sunderland, Mass: Sinauer As
REFERENCE BOOKS:
1. Lewin B.. (2000) Gene VIL IRL Press, Oxford University Press Oxford.
2. Watson, J.D., Hopkins, Roberts, Stiez, Weiner. (1987) Molecular Biology of the Gene. (4th ed) The Benjamin/Cummings Publishing Co. Inc.
California.
3. Davis. D.B.. Dulbecco, R., Risen, H.N, Ginsberg. H.S., (1990) Microbiology, (4th ed) harper &Row Publishers, Singapore.
4. Principles of biochemistry, Lehninger, A.L. 2nd ed 1993
5. Biotechnology, Smith John. 3rd ed. 1996
6. Pelczar, Jr. Ml, Chan, E.C.S.. Krieg. N.R. 11986) Microbiology (5th ed). McGraw-Hill Book Company. NY.
7. Madigan, M.T., Martinko, J.M.. and Parker, J. ('2003) Brock Biology of Microorganisms. (10th ed). Prentice-Hall International N.I USA.
8. Williams. B.L. and Wilson, K.., (ed). (1981) A Biologist's Guide to Principles and Techniques of Practical Biochemistry, (2nd ed). Edward Arnold
London.
(A) Isolation and identification of the following medically important bacteria.
- E. coli - Enterobactor aerogenes
- Proteus vulgaris - Salmonella typhi
- Salmonella paratyphi A - Salmonella paratyphi A
- Shigella dysentriae - Pseudomonas aeroginosa
- Staphylococcus aureus
(B) Preparation of serum
(C) study of serological reactions
(i) Agglutination reaction
(ii) Precipitation reaction.
(D) Study of permanent slides of medically importance bacteria, vectors and plant pathogens.
(i) Pneumococci, Spirochetes
(ii) Tick, mite, Flea, female anopheles, Bed bug, Head louse, Body louse.
(iii) Paccinia graminis :Uredospore, Teleospore, aciospore, pycniospore stages.
(E) Study of Plant disease
Downy mildew of bajaro
Powdery mildew of grapes
Smut of wheat/ jowar
Rust of wheat
Citrus cancer
Angular leaf spot of cotton
Tobacco mosaic disease.
Microbial technology
(A) Isolation and identification and study of morphological characteristics of various microorganisms of biological interest:
(i) Bacillus
Bacillus subtillis, B. megaterian, B. cereus, Actinomycetes, Azotobacter, Rhizobium, kanthomonas.
(ii) Yeast and Mold
Yeast, Rhizopus, Mucor, Aspergillus, Penicilium, Neurospora, Fusariu'm
(B) Hematology
(i) Total count of RBCs and WBCs
(ii) Hemoglobin estimation
(iii) ESR
(iv) Differential count of WBCs
(v) Blood grouping
(C) Clinical biochemistry :
(i) Estimation of blood sugar by glucose oxidase method.Total count of RBCs and WBCs
(ii) Estimation of blood urea by diacetyl monoxime method.
(D) Urine Analysis
Physical and Chemical analysis Microscopic examination.
(E) Diagnostic microbiology:
Identification of unknown bacteria from mixed population using identification key
(F) Serodiagnosis of diseases:
(i) Serological diagnosis of entric fever using Drayer' method and double dilution method.
(ii) Serological diagnosis of syphilis using VDRL test.
(5) Industrial Microbiology
(a) Primary screening of important microorganisms capable of producing bioactive substances
i. Screening of antibiotic producers
ii. Screening of organic acids producers
iii. Screening of amylase producers
(b) Bioassay of penicillin / cephalosporin using Bacillus subtillis. Primary screening of important microorganisms capable of producing
bioactive substances
(c) Sterility testing of pharmaceutical products
(d) Study of enzyme (amylase) activity
(e) Fermentative production amylase
Visits to public Health or Drug Laboratories and institutions will be undertaken during the year.
The candidate is required to bring for inspection at the practical examination his/her own practical journal duly-signed and certified by the Head of the Department.
The oral examination will be illustrative of the theoretical portions of the syllabus.