DEPARTMENT OF BIOTECHNOLOGY - Bundelkhand … · department of biotechnology bundelkhand...

DEPARTMENT OF BIOTECHNOLOGY

BUNDELKHAND UNIVERSITY, JHANSI

COURSE CURRICULUM

M. Sc. BIOTECHNOLOGY

Semester Paper

Code TITLE OF THE THEORY / PRACTICAL PAPER SA IA

Total

marks

6001

(BT-101) BIOCHEMISTRY 70 30 100

6002

(BT-102) MOLECULAR CELL BIOLOGY AND GENETICS 70 30 100

6003

(BT-103) BIOINFORMATICS AND BIOSTATISTICS 70 30 100

6004

(BT-104) CHEMISTRY OF BIOMOLECULES 70 30 100

6005

(BT-105) BIOPHYSICAL CHEMISTRY AND INSTRUMENTATION 70 30 100

1st

6006

(BT-106) LAB TECHNIQUES-I BIOCHEMISTRY 100 --- 100

6007

(BT-201) ENZYME TECHNOLOGY 70 30 100

6008

(BT-202) IMMUNOTECHNOLOGY 70 30 100

6009

(BT-203) MOLECULAR BIOLOGY 70 30 100

6010

(BT-204) MICROBIOLOGY 70 30 100

6011

(BT-205) BIODIVERSITY, BIOETHICS & IPR 70 30 100

2nd

6012

(BT-206)

LAB TECHNIQUES-II MICROBIOLOGY AND

IMMUNOLOGY 100 --- 100

7001

(BT-301) INDUSTRIAL BIOTECHNOLOGY 70 30 100

7002

(BT-302) GENETIC ENGINEERING AND ITS APPLICATIONS 70 30 100

7003

(BT-303) ANIMAL BIOTECHNOLOGY 70 30 100

7004(BT

-304) PLANT BIOTECHNOLOGY 70 30 100

7005

(BT-305) LAB TECHNIQUES-III (A) MOLECULAR BIOLOGY 100 --- 100

3rd

7006

(BT-306) LAB TECHNIQUES-III (B) INDUSTRIAL TECHNOLOGY 100 --- 100

4th

7007

(BT-401) PROJECT / DISSERTATION 200 --- 200

GRANT TOTAL MARKS 2000



6001 (BT-101) BIOCHEMISTRY

UNIT I Carbohydrate Metabolism

Basic concepts, Glycogenesis, gluconeogenesis : pathway and regulation, Glycolysis, krebs cycle, pentose

phosphate pathway, glyoxalate pathway, glycogenolysis : pathway and regulation

UNIT II Electron transport chain and Oxidative phosphorylation

Electron carriers, Complexes I to IV, Chemiosmotic theory, Substrate level phosphorylation

UNIT III Lipid Metabolism

Biosynthesis and degradation of odd and even carbon: saturated and unsaturated fatty acids. Formation of

Ketone bodies, Regulation of metabolism, associated inborn errors

UNIT IV Amino Acid metabolism

Amino acid oxidation and production of urea. Metabolic fates of amino groups. Nitrogen excretion and

urea cycle. Overview of nitrogen metabolism.

UNIT V Nucleic acid Metabolism

Biosynthesis and breakdown of purine and pyrimidine nucleotides by denovo and salvage pathway.

UNIT VI Integration of metabolism

Role of Liver, Adipose tissue, Muscle, and Brain, Role of blood as carrier of O2, metabolites and hormones.

Hormonal regulation of fuel metabolism.

List of suggested reading:

1. Lehninger Principles of Biochemistry. Nelson, D.L and Cox, M.M.

2. Biochemistry Stryer

3. Biochemistry . Zubay,G

4. Fundamentals of Biochemistry . Jain, J.L.

5. Fundamentals of Biochemistry. Voet, D and Voet, J.G.

6. Biochemistry . Freifilder, D.

7. Fundamentals of Biochemistry . Deb, A.C.



6002 (BT- 102) MOLECULAR CELL BIOLOGY AND GENETICS

PART A. MOLECULAR CELL BIOLOGY

UNIT Basics of Cell Biology

Definition of cell- fundamental cell types, difference between Prokaryotic and Eukaryotic cell types,

Structure and function of cell organelles, ultra structure of cell membrane, cytosol, Golgi bodies,

endoplasmic reticulum (rough and smooth), ribosomes, cytoskeletal structures (actin, microtubules etc.)

Mitochondria, chloroplasts, lysosomes, peroxysomes, nucleus (nuclear membrane, nucleoplasm, nucleolus

chromatin).

UNIT II Biomembranes

Structure and functions, Biological membranes and transport of solutes. Molecular structure of membrane.

Solute transport across membranes. Types of transport. Simple, passive, facilitated diffusion. Active

transport. Transport ATPases. Mobile carriers and pore mechanism. Artificial membranes. Micells, bilayer,

monolayer, liposomes.

UNIT III Cell signaling and signal transduction

Types of cellular receptors and chemical signals. G proteins. Second messengers. Role of steroid hormones.

UNIT IV Cell cycle

Cell division and cell cycle: mitosis, meiosis, stages of cell cycle, binary fission amitosis.

Cell-cell interaction, Cell senescence and death: Apoptosis and necrosis

PART B. GENETICS

UNIT I History

Gene organization and expression in prokaryotes and eukaryotes. Mendelian Laws of inheritance, gene

interactions.

Extrachromosomal inheritance, mitochondrial and chloroplast genetic systems: sex linked inheritance.

UNIT II Linkage and crossing over

Sex linked inheritance, Maternal effects and cytoplasmic inheritance, Structural and numerical changes in

chromosomes, Hereditary defects. Inbreeding depression and heterosis

UNIT III Mutations

Mutation: Spontaneous versus induced mutations, types of mutations, the molecular basis of mutations,

mechanisms of DNA repair, mutations, frequency, correlation between mutagenicity and carcinogenicity,

mutagenic agents, chemical and radiation. Population Genetics: Hardy-Weinberg equilibrium, gene and

genotypic frequencies, introduction of eugenics.


1. Molecular Biology of the cell. Alberts, B. et al

2. Molecular Cell Biology. Lodish et al,

3. Cell Biology. De Robertes.

4. Principles of Genetics. Gardener, Simmons and Snustad



5. Cell and Molecular Biology. Gupta, P.K.



6003 (BT-103) BIOINFORMATICS AND BIOSTATISTICS

A. COMPUTER FUNDAMENTALS

UNIT I Introduction

Hardware, storage and memory devices, input and output devices, file and folder concepts, operating

systems, file management, network, internet and e-mail. Printers, Floppies, Mouse, Key board, Booting, MS

Office, Windows.

UNIT II Net working

LAN, MAN, WAN, WWW, HTML, programming language.

UNIT III Application software

Introduction to office suit, utility and capabilities of word: processing, spreadsheet, database and

presentation utilities

B. BIOINFORMATICS

UNIT I

History, scope and applications of Bioinformatics, database

UNIT II

DNA Sequence Information Sources, EMBL, GENBANK, DDBJ, Entrez, Unigene, Understanding the

structure of each source and using it on the web.

Protein Information Sources, PIR, PDB, SWISSPROT, TREMBL, Understanding the structure of each source

and using it on the web.

Biological metabolic pathway database- KEGG

Introduction of Data Generating Techniques, Restriction Enzymes, Gel Electrophoresis, Chromatograms,

Blots, PCR, Microarrays, Mass Spectrometry, What data each generates and what Bioinformatics problems

they pose

UNIT III

Sequence and Phylogeny analysis, Detecting Open Reading Frames, sequence alignments, Introduction to

BLAST, using it on the web, Interpreting results, Multiple Sequence Alignment Phylogenetic Analysis.

Searching Databases: SRS, Entrez, Sequence Similarity Searches-BLAST, FASTA and DATA Submission

Protein Structure: Protein structure classification, Structure Analysis, Secondary structure prediction

methods, Ramachandran plot, homology modeling

Drug discovery and pharmcogenomics

Genome Annotation: Pattern and repeat finding, Gene identification tools.



C. BIOSTATISTICS

UNIT I Introduction

Concept of variables in biological systems, Collection, classification, tabulation, graphical and diagrammatic

representation of numerical data

UNIT II Measures of central tendency

Mean, median and mode and their relationship, HM, Quartices

UNIT III Measures of Dispersion

Range Mean deviation, Mean absolute Deviation, Mean squared deviation (variance), Root-Mean Square

deviation (standard deviation) Coefficient of variation, Quartice deviation.

UNIT IV Probabilty and Distributions

Introduction, Random experiment, Sample space, Events, Addition and multiplication theorem of

probability, Conditional probability, Bayes theorem, Random variables and Probability distributions.

Binomial, Poisson distribution, Applications of probability and distributions.

UNIT V Tests of Significance

Based on T, F, Z and Chi-square distributions.

UNIT VI Analysis of Variance

One-way and Two way classifications.

UNIT VII Correlation and Regression

Correlation Coefficient, Rank correlation. Regression lines (YonX-Xon Y). Regression coefficient.

Principles of experimental design. Completely randomixed, randomized block. Latin sequence design.

ANOVA. Statistical and graphic software-their use in research for data analysis, interpretation and

presentation.


1. Bioinformatics. Baxevanis, A.D. and Quelette, B.F.F.

2. Bio informatics. Des Higgins & Willie Taylor

3. Bioinformatics. Methods and protocols. Macsewer, S.

4. Bioinformatics. Sequence and genome analysis. Mount, D.W.

5. Computer fundamentals. Nagpal, D.P.

6. Methods in Biostatics. Mahajan,B.K.

7. Biostatistics. Arora, P.N. and Malhan, P.K.

8. Elements of Biostatistics .Prasad, S.

9. Practical in statistics. Sharma, H.L.

10. Fundamentals of Mathematical Statistics. Gupta, S.C. and Kapoor, V.K.

11. Mathematics for Biosciences - Arya J.C and Lardner, R.W

12. Advanced Engineering Mathematics-Erwin Kreyszig



13. Biostatics -P.N. Arora & P.K. Malhotra

14. Introduction to Biostatistics- Sokal & Rohif

15. Statistical Methods in Bioinformatics: An introduction-W.Evans.Grant

16. Computers Today-S.K .Basandra

17. Computer fundamentals-P. K. Sinha



6004 (BT-104) CHEMISTRY OF BIOMOLECULES

UNIT I Atom and Chemical Bonds

Structure of an Atom. Ionization potential. Covalent, Ionic, Coordinate and Hydrogen bonds. Disulphide

bonds. Vanderwaals interactions,Hydrophobic interactions. Electronegativity, Polar and non polar covalent

bonds. Bond length and angle, Bond energy, Strength of bonds in common biomolecules

UNIT II Chemistry of nucleic acids

Structure of bases and sugars involved. Tautomerism of bases. Chargaffs rules. Nucleotides. Base

modifications. Double helix (Watson and Crick model). Various non B forms and their structural features.

Evidence for the presence of hydrogen bonds and hydrophobic interactions in DNA. Palindromes. Hair pin

and cruciforms. Banding of DNA and its applications. Supercoiling. Linking number. Enzymes involved in

supercoiling. Toroidal and solenoid forms. DNA packing in chromatin. Multiplex structure of DNA and its

relevance. Triplex. H-DNA, Tetraplexes. Telomeres-structure and functions. DNA denaturation and

renaturation, Cot values, Buoyant density, c-value.

UNIT III Chemistry of Carbohydrates

Classification of Carbohydrates. Biological significance. Structure of glucose, fructose, mannose, galactose,

lactose, sucrose, glycogen, starch and cellulose. Chemical properties of sugars. Isomerism, Structural and

sterioisomers- optical, geometric. Mutarotation, Enantiomers, epimers, and Diasterioisomers. Anomers.

UNIT IV Chemistry of Lipids

Classification and biological sygnifcance Triglycerides, Waxes Phosphopids, sphingolipids, Sterols,

Lipoproteins, Steroid hormones and their functions. Cholesterol and prostaglandins, Physiological

significance. Fats and Oils, Classification of fatty acids, essential fatty acids.

UNIT V Chemistry of proteins

Classification of proteins - functional, composition and solubility, Amino acids, Classification and structure.

Essential amino acids, Non protein and non standard amino acids, Physical and chemical properties of

proteins. Reactions involving, NH2 Group, COOH and R Groups.

UNIT VI pH and Buffers

Ionization of water, weak acids and bases. The pH scale, The bronsted lowry concept of acids and bases.

Titration of weak acid by a strong base, Henderson-Hasselbalch equation, Biological buffer systems. The

bicarbonate, amino acid, hemoglobin and protein buffer systems. Acidosis and alkalosis


1. Fundamentals of Biochemistry. Jain, J.L.

2. A Text Book of Biochemistry. Singh, S.P.

3. Biochemistry. Stryer L.

4. Lehninger Principles of Biochemistry. Nelson, D.L. and Cox, M.M.

5. The Biochemistry of Nucleic acids. Adams et al

6. Fundamentals of Biochemistry. Deb, A.C.

7. Biochemistry- Rawn, J.D.



8. Principles of Biochemistry- Lehninger, A.L. Nelson, D.L. and Cox, M.M

9. Principles of Biochemistry: Student Study Art Notebook- Zubay, G.L, Parson, W.W. and Vance, D.E.

10. Carbohydrate Biotechnology Protocols- Bucke C.

11. Principles of Biochemistry- Horton et al.

12. An Introduction of practical biochemistry- Plummer D.T.

13. Practical Biochemistry- Bansal, D.D., Khardori, R & Gupta, M.M.



6005 (BT-105) BIOPHYSICAL CHEMISTRY AND INSTRUMENTATION

UNIT I Microscopy

Principles and applications of light microscope, fluorescence, Phase contrast, Dark field, electron (TEM and

SEM), interference, polarization, inverted and stereomicroscope. Freeze fracture fixation and staining.

UNIT II Spectrophotometry

Photometry - principles of UV-Vis spectrophotometry, instrumentation and applications, Fluorimetry -

principle instrumentation and applications, X-ray Crystallography - principle, instrumentation and

applications, Flame techniques, atomic absorption and flame emission spectroscopy-principles,

instrumentation and applications, NMR - principle, instrumentation and applications, ESR - principle,

instrumentation and applications

UNIT III Radiation Biology

Radioisotope Techniques: Radio-tracers, interaction of radiation with matter, GM counter, Proportional and

Scintillation counters, methods of quench correction autoradiography, radioimmunoassay.

UNIT IV Chromatographic Techniques

Principle, types and applications- partition, adsorption, gel filtration, paper, thin layer, gas

chromatography, ion exchange, molecular sieve HPLC - principle, instrumentation and applications.

UNIT V Centrifugation

Principle, types: differential density gradient centrifugation, sedimentation coefficient, factors affecting

sedimentation coefficient and applications.

UNIT VI Electrophoresis Techniques

Principle types and applications: polyacrylamide and agarose gel electrophoresis. Isoelectric focusing.


1. Physical Biochemistry: Applications to Biochemistry and Molecular Biology. Freifelder, D.

2. Principles of Instrumental Analysis. Skoog and West

3. A Biologists Guide to Principles and Techniques of Practical Biochemistry. Wilson, K. and Goulding, K.H.

4. Principles of Instrumental Analysis. Skoog, Hollar and Nieman

5. Biological Spectroscopy. Campbell and Dwek

6. Introductory practical Biochemistry. Sawhney, S. K and Singh, R.

7. Bioseparations: Principles and Techniques. Sivasankar. B. 2007

8. Principles and techniques of practical Biochemistry. Eds K. Wilson and J. Walker



6006 (BT-106) LAB TECHNIQUES -I

1. Preparation of buffers (Citrate, phosphate, tris and acetate)

2. Proteins

a) Extraction of proteins

b) Wavelength scans of proteins

c) Estimation of proteins by Lowry and Bradford method

3. Nucleic acids

a) Estimation of DNA and RNA

b) Melting studies of DNA

4. Carbohydrates

a) Estimation of total carbohydrate content of sample by anthrone method

b) Colour reactions for identification of carbohydrates

c) Estimation of starch by anthrone reagent

d) Extraction and estimation of chlorophyll

5. Amino Acids

a) Estimation of total free amino acids

b) Chromatography- TLC and paper


1. Biochemical calculations Segal, I.H.

2. Biochemical Methods Sadashivam, S and Manikam, A.

3. A Biologists guide to principles and practices in Biochemistry. Wilson, K. and Goulding, K.H.

4. Bioseparations-Principles and Techniques. Sivasankar B. 2007



6007 (BT-201) ENZYME TECHNOLOGY

UNIT I Introduction

History, general characteristics, nomenclature, IUB enzyme classification. Holoenzyme, apoenzymes,

coenzymes, cofactors. Activators and inhibitors. Metalloenzymes. Units of enzyme activity. Isoenzymes.

Monomeric and oligomeric enzymes. Multienzyme complexes. I.U. Katal, turnover number and specific

activity. Role of vitamins as coenzymes precursors. Enzyme catalysis. Acid base, covalent. Proximity and

orientation effects.

UNIT II Specificity and models of enzyme action

Substrate and reaction specificity. Lock and key hypothesis. Induced fit hypothesis. Strain and transition

state hypothesis.

UNIT III Enzyme kinetics

Factors affecting enzyme activity. Enzyme concentration, substrate concentration, pH and temperature.

Michaelis-Menton equation for unisubstrate reactions. Km and its significance. Briggs Haldane steady

state. Line weaver-Burk plot and its limitations. Sequential and ping pong mechanisms. Kinetics of Zero and

first order reactions. Enzyme inhibition- reversible and irreversible. Competitive, noncompetitive and

uncompetitive inhibitors. Determination of km and V max in presence of inhibitors.

UNIT IV Allosteric enzymes

Sigmoidal kinetics. Symmetric and sequential model for action of allosteric and their Significance. Negative

and positive cooperativity and half side reactivity. Effect of partition on kinetics and performance with

particular emphasis on changes in pH and hydrophobicity.

UNIT V Mechanism of enzyme regulation

Feedback inhibition and feedforwad stimulation. Enzyme repression, induction and degradation. Control of

enzymatic activity by product and substrates. Reversible and irriversble covalent modification of enzymes.

Monocyclic and multicyclic cascade system. Mechanism of activation of Zymogens.

UNIT VI Enzyme Technology

Isolation and purification and crystallization of enzymes. Applications of enzymes in industries and

medicine. Production of immobilized enzyme and their applications. Engineering of enzymes by site

directed mutagenesis. Active site mapping. Nature of active site. Enzyme assays for diagnosis and

prognosis of diseases. Enzyme electrodes. Enzyme immunoassay. Enzymes from extreme environments.


1. Understanding Enzymes. Palmer,T. 2. Enzyme. Dixon, M. and Webb, E.C.

3. Fundamentals of Biochemistry. Jain, J.L. 4. Fundamentals of Biochemistry. A.C.

5. General Enzymology. Kulkarni and Deshpande 6. Enzyme reaction Mechanisms. Welsch.

7. Lehninger Principles of Biochemistry. Nelson and Cox

8. Enzyme Kinetics. A modern approach. Marangoni, A.G.

9 .Fundamentals of Enzymology. Price, N.C. and Stevens, L.



10. Enzyme Assays: A practical approach. Eisenthal and Danson



6008 (BT-202) IMMUNOTECHNOLOGY

UNIT I Immune system

Immune response - Innate and Adaptive Immunity and Characteristics, Anatomical organization of immune system –

Primary and secondary lymphoid organs, Secondary lymphoid organs, Cells of the immune system, lymphocyte

traffic. Hematopioesis, Antigens Characters, Haptens, Antigenicity and Immunogenicity, Factors affecting

immunogenecity, Properties of T and B cell epitopes, Super antigens. Immunoglobulins – structure and functions,

Organisation and expression of immunoglobulin genes. Mechanisms of antibody diversity, Class switching,

Immunoglobulin Superfamily Complement System Components, activation pathways and their regulation,

Complement deficiencies and role of complement system in immune response

UNIT II Major Histocompatibility systems

Organization of MHC I and II molecules, polymorphism, distribution and function, organization of MHC complex in

Mouse and Humans. MHC and disease resistance T-Cell receptor – Structure and function, T- cell accessory

membrane molecules, activation of T-cells B-Cell receptor –Structure, types and functions, activation of B-cells

UNIT III Immune effecter mechanisms

Structure and functions, cytokine receptors, signal transduction mediated by cytokine receptors, cytokine related

diseases and their therapeutic applications Antigen presentation – Processing and presentation of exogenous and

endogenous antigens. Immunoregulation – mediated by antigens, antibodies, immune complexes. Complement

System, Components, activation pathways and their regulation, Complement deficiencies and role of complement

system in immune response, Hypersensitivity – Definition, Type I, II, III and IV type hypersensitivity, delayed type of

hypersensitivity, Autoimmunity – Organ specific and systemic diseases, mechanisms of autoimmunity and

therapeutic approaches

UNIT IV Role of immune system in Health and Diseases

Immunodeficiency Syndromes – Primary and secondary immunodeficiencies, their diagnosis and therapeutic

approach, AIDS

UNIT V Immunity and Biotechnology

Antigen and Antibody interactions – Affinity, Avidity, Cross reactivity, forces involved in Ag-Ab interaction, Methods

of immunodiagnosis. Antibody engineering. Expression systems for antibody formation. In vitro libraries for

antibody identification, Chimeric antibodies. Vaccines – Active and passive immunization, whole organism vaccines,

Macromolecules as vaccines, Recombinant-vector vaccines, DNA vaccines, synthetic peptide vaccines and subunit

vaccines.

UNIT VI Immunological Techniques in Biotechnology

Hybridoma technology and production of monoclonal antibodies. Culture of hybridoma cells. Purification of

antibodies. Characterization and applications in diagnosis, therapy and basic research.


1. Immunology Roitt, Brostoff and Male 2. Essential Immunology. Roitt, Ivan.M.

3. Fundamental of Immunology (2008). William Paul

4. Immunology Goldsby, R.A., Thomas J Kindt, Barbara, A Osborne and Kuby, Janis. W.H.

5. Immunology: An introduction. Ian R. Tizard



6009 (BT-203) MOLECULAR BIOLOGY

UNIT 1 Introduction

History, Nucleic acids as genetic material, Transformation Hershey and Chase experiments. TMV

reconstitution experiment. Central dogma of molecular genetics. Current version. Reverse transcription.

UNIT II Replication

General principles. Replicon. Origin of replication. Bidirectional and Unidirectional replication. John Cairns

experiment. Semiconservative replication. Messelson and Stahl experiment. Inmans experiment on

replication of DNA.(J.M.B.1970. 51: 61). Primase and primasomes. Origin of replication. Role of RNAase H.

(a) Prokaryotic replication

Functions and properties of enzymes involved in replication. DNA polymerase I, II and III. Structure and

functions. DNA Ligase. Helicases and topoisomerases. Single strand binding proteins. Initiation and

termination. Plasmid DNA replication. Rolling circle model of replication x 174 RNA replication in RNA

viruses. RNA Replicase. Reverse transcription. Replication in phage 29 and adenoviruses.

(b) Eukaryotic Replication

Eukaryotic DNA Polymerases. Location and functions. End completion of linear DNA. Synthesis of telomere

DNA. Termination in T7 phage. Concatamers. Huberman and Riggs experiment ( J.M.B. 1968. 32: 327.) D

loops. Mitochondrial and chloroplast DNA replication.

UNIT III Transcription

(a) Prokaryotic transcription: Promoters, Constitutive and inducible, Operators. RNA polymerase. Subunit

structure . Role of different sigma factors in differential expression of genes in bacteria. Initiation,

elongation and termination. dependent and independent termination. Inhibitors of prokaryotic

transcription. Attenuation ,antitermination.

(b) Eukaryotic transcription: Eukaryotic promoters and enhancers, Location of promoter sequences.

Eukaryotic RNA polymerases I, II and III and their products. Initiation by polymerases I, II and III.

Transcription factors. Cis-acting and trans acting factors. TATA binding proteins. Inhibitors of transcription.

Regulation of transcription by steroid hormones. Prokaryotic and Eukaryotic mRNA. Life span and

degradation.

UNIT IV Translation

Genetic code-salient features. Altered genetic code in mitochondria. Prokaryotic and eukarytic ribosomes.

rRNA and tRNA. Structure of yeast phe. tRNA. Nonsense, missense and suppressor mutations. Process of

translation. Activation of aminoacids. Amino acyl tRNA synthetases Initiation, Elongation, Translocation

and Termination in proparyotes and eukaryotes. Initiation and elongation factors. Inhibitors of translation.

Translation in chloroplasts and mitochondria.

UNIT V Protein targeting and degradation

Modification, folding and transport of polypeptides. Chaperones. Control of cellular location of the

products of translation. Regulation of protein synthesis. Signal hypothesis, glycosylation of proteins in E.R.

Protein transport across membranes. Transport to mitochondria, lysosomes, chloroplasts, nucleus and



peroxisomes. Polyprotein precursors of peptide hormones. Protein degradation in prokaryotes and

eukaryotes. Proteosomes. ubiquitin.

UNIT VI Modification of nucleic acids

RNA and DNA methylation. Role of methylation in protecting from restriction endonucleases. Methylases

and their use in creating extended restriction enzyme cleavage sites. Demethylation and gene expression.

UNIT VII DNA repair

Pyrimidine dimers. Repair mechanisms. Photoreactivation. Excision- photolyase, methyl transferases,

glycosylation. A.P. site. Mismatch, Post replication repair. Recombination repair. SOS repair.


1. The Biochemistry of nucleic acids. Adams et al.

2. Molecular Biology of the gene. Watson et al.

3. Lehninger Principles of Biochemistry. Nelson and Cox.

4. Genes VIII. Lewin, B.

5. Principles of Genetics. Gardener, Simmons and Snustard.

6. Molecular Biology and Genetic engineering. Gupta, P.K.



6010 (BT-204) MICROBIOLOGY

UNIT I Introduction

History of Microbiology, Bacteria, Cyanobacteria and fungal cell. Bacteria-size, shape, capsules, gram

negative and gram positive cell wall, structures external to cell wall, Concept of microbial growth, effect of

environmental factors on growth such as salt concentration, pH, temperature etc., growth monitoring and

characteristics. Concept of nutrition (micro, macro and trace nutrients), types of nutrition (passive, facilitated,

and active transportation) and categories of microorganisms on the basis of mode of nutrition.

UNIT II Basic concepts

Methods of classification and major groups of microorganisms. Taxonomic nomenclature and Bergey’s

Manual. Concept of sterilization in microbiology

UNIT III Pathogenicity of microorganisms

Bacterial and Viral pathogenesis, Microbial mechanisms for escaping host defenses. Endospore formation.

sanitary analysis of water

UNIT IV Microbial Metabolism

Fermentation and its products, peptidoglycan synthesis and LPS synthesis

UNIT V Microbial genetics

Bacterial Plasmids-Characters, copy number, compatibility, classification (F, R, Col, degradative, heavy

metal resistance, Ti, cryptic), yeast plasmid. Microbial genetics and recombination: mutations, mutant

screening, enrichment and selection, conjugation, transduction and transformation.

UNIT VI Viruses

Virology: Overview of virus, classification of animal and plant viruses, types of viral replication, Titration of

viruses, General structure and life cycle of bacteriophage, Hepatic virus (HAV, HAB, HAC), Dengue virus (Flavi

virus), Bird flu virus (Avian influenza virus)

UNIT VII Chemothearpy and antibiotics

Antimicrobial agents, sulpha drugs, antifungals, mode of action and resistance to antibiotics, Antibiotic

sensitivity assays


1. Microbiology. Prescott, L.M., Harley, J.P and Klein, D.A.

2. Brock Biology of Microorganisms. Martinko. C.M.T. and Parker, J.M

3. Microbiology. Pelczar, C.M.J., Chan, E.C.S and Krieg, N.R.

4. General Microbiology. Stanier, C.

5. Microbiology-a laboratory manual Cappuccino, J.G. and Sherman, N. and Weseleey, Addison.

6. Experiments in Microbiology, Plant Pathology and Biotechnology. Aneja, A.R.

7. Microbial metabolism. Doelle.

8. Microbiology an Introduction. Tortora.

9. Microbiology Fundamentals and Applications. Atlas, R.M.



6011 (BT-205) BIODIVERSITY, BIOETHICS & IPR

UNIT I Introduction to biodiversity

Levels of biodiversity -values of biodiversity - loss of biodiversity - Species concept - Classification and

systematic: biological nomenclature - biological classification; Biodiversity conservation: in situ and ex situ -

Magnitude and distribution of biodiversity - wild life biology - conservation strategies - measures of

biodiversity - biodiversity in India and global level - biodiversity hot spots.

UNIT II Introduction to ethics/bioethics

Framework for ethical decision making; biotechnology and ethics - biotechnology in agriculture and

environment: benefits and risks – benefits and risks of genetic engineering - ethical aspects of genetic

testing - ethical aspects relating to use of genetic information - genetic engineering and biowarfare.

UNIT III Ethical implications of cloning

Reproductive cloning , therapeutic cloning ; Ethical, legal and socio-economic aspects of gene therapy,

germ line, somatic, embryonic and adult stem cell research- GM crops and GMO's - biotechnology and

biopiracy - ELSI of human genome project.

UNIT IV Introduction to Biosafety

biosafety issues in biotechnology - risk assessment and risk management - safety protocols: risk groups -

biosafety levels - biosafety guidelines and regulations (National and International) - operation of biosafety

guidelines and regulations - types of biosafety containments.

UNIT V Introduction to intellectual property and intellectual property rights

Types: patents, copy rights, trade marks, design rights, geographical indications - importance of IPR

patentable and non patentable - patenting life - legal protection of biotechnological inventions - world

intellectual property rights organization (WIPO)

Entrepreneurship

Selection of a product, line design and development processes, economics on material and energy

requirement, stock the product and release the same for making etc. The basic regulations of excise:

Demand for a given product, feasibility of its production under given constraints of raw material, energy

input, financial situations export potential etc.


1. S. Ignachimuthu,S.J., Bioethics, Narosa Book Distributors Pvt. Ltd., Delhi, 2009

2. M.K. Sateesh, Bioethics and Biosafety, I. K. International, Pub. House Pvt.Ltd., New Delhi, 2008

3. Ben Mepham, Bioethics, II Edn., Oxford University Press, Hampshire, Great Britain, 2008

4. J.E. Smith (2004). Biotechnology (Cambridge Univ. Press).

5. V. Santaniello (2000). Agriculture and intellectual property rights: economic institutional, and

implementation issues in biotechnology (Edition illustrated Publisher CABI).

6. Thackerey, A (ed) (1998). Private science: Biotechnology and the rise of the molecular sciences (Univ. of

Pennsylvania Press, Phil).



6012 (BT-206) LAB TECHNIQUES-II

A. MICROBIOLOGY

1. To study the available laboratory equipments.

2. To prepare media for routine cultivation of bacteria (Nutrient Agar; NA) and fungi (Potato Dextrose Agar; PDA and

Czapek’s Dox agar; CDA)

3. To prepare agar plates, agar slants and to practice streaking for isolated colonies.

4. To observe given fungal cultures by lacto phenol cotton blue mounting technique (Mucor, Alternaria, Penicillium,

Rhizopus and Aspergillus).

5. To observe and count the microorganisms present the given milk sample by standard Plate count method.

6. a) To observe and count the microorganisms present the given soil (non-rhizosphere) sample by standard Plate count

method.

b) To observe and count the microorganisms present the given soil (rhizosphere) sample by standard Plate count

method and calculate the R:S values.

7. To analyze the quality of milk sample by methylene blue reductase test.

8. To observe the bacterial cell arrangement pattern by negative staining.

9. To observe the arrangement pattern and shape of bacterial cells by gram staining.

10. To test the antibiotic sensitivity of given cultures by disk diffusion method (Stoke’s method).

11. To perform E-test for determining MIC of drug on the given culture.

12. To observe the sigmoid growth pattern of given bacterial culture.

13. To prepare differential agar plates (Mannitol agar) and to observe mannitol fermenting and non-fermenting colonies on

the plate

14. To perform various biochemical tests for the biotyping given bacterial cultures

a) Colony characterization

b) Gram staining (Differential gram stain)

c) Motiliy test (Nutrient deep agar tubes)

d) Catalase test (3% H2O2)

e) Oxidase test (DMPD)

f) Citrate utilization (Simmon’s Citrate Agar)

g) Urease test (Christensen’s media)

h) MR-VP test ( MR-VP broth)

i) H2S production test (SIM deep agar tubes)

j) Carbohydrate fermentation-mannitol, mannose, xylose, arabinose, glucose, fructose, dulcitol, rhamnose (Peptone

water)

k) Indole production test

l) TSI test (Triple Sugar Iron test)

B. IMMUNOLOGY

Practicals will be performed on the type of availability of teaching kits.


1. Microbiology. A Laboratory manual. Cappucino, J.G., Sherman, N. and Weseeley, A.

2. Experiments in Microbiology, Plant Pathology and Biotechnology. Aneja, A.R.

3. Microbes in Action.



7001 (BT-301) INDUSTRIAL BIOTECHNOLOGY

UNIT I Introduction

Definition, Scope and chronological development of the fermentation industry, the component parts of a

fermentation process.

UNIT II Improvement of industrial micro-organisms

Isolation of industrially important micro-organisms – Screening, selection of mutants overproducing

primary metabolites, selection of mutants overproducing secondary metabolites, use of recombination

systems for the improvement of industrial micro-organisms and by modifying properties other than the

yield of product. Maintenance and preservation of microorganisms

UNIT III Media design

Introduction, typical media - energy sources, carbon and nitrogen sources and other components

UNIT IV Sterilization

Medium sterilization, design of a batch and continuous sterilization processes, holding time, Scale up of

sterilization.

UNIT V Fermenter design

Basic concepts of fermenter-batch,continous, fed batch, packed bed, bubble column, fludised bed, trickle,

plug flow reactors. Fermentations-Submerged and solid state, body construction, aeration and agitation.

UNIT VI Recovery and purification of fermentation products

Removal of microbial cells and other solid matter, foam separation, precipitation, filtration, centrifugation,

cell disruption, Liquid- liquid extraction, drying and crystallization.

UNIT VII Microbial production

Penicillin, Riboflavin, citric acid, vinegar, beer and whisky, Glutamic acid, Bread and Mushroom production

UNIT VIII Effluent treatment

Treatment and Disposal of effluents, B.O.D. and C.O.D.


1. Biochemical Reactors Atkinson, B. Pion Ltd London

2. Biochemical Engineering Fundamentals Baily, J.F. and D F Ollis, D.F.

3. Bioprocess Technology-Fundamentals and Applications KTH Stockholm

4. Principles of fermentation technology Stanbury, P.F. and Whitakar, A.

5. Bioprocess Engineering principles, Pauline M. Doran, Academic Press



7002 (BT-302) GENETIC ENGINEERING AND ITS APPLICATIONS

UNIT I Tools for Recombinant DNA Technology

Nucleases and restriction endonucleases – properties and types : phosphomonoesterases, polynucleotide

kinase, DNA ligase, DNA polymerase I, RNA dependent DNA polymerase, terminal deoxynucleotidyl

transferase, Poly A polymerase

UNIT II Cloning Vectors

Prokaryotic host – vector system – Characteristics of E. coli as host, vectors for cloning in E. coli (plasmid,

bacteriophage, cosmid) Plasmid cloning vectors. Size, range, phenotypic traits, cryptic and conjugative/no

conjugative. Stringent relaxed. Phages as cloning vectors. Advantages in the use of phage lamda and other

usefull phages. M13 and its use in DNA sequencing. Cosmid vectors. Construction of cosmid DNA and its

advantages. Phasmid vectors. Yeast vectors. Vectors for plants and animals. (Ti plasmids and viral vectors-

caulimovirus, geminivirus) and animal system (SV 40, bovine papilloma virus, adenovius, vaccinia virus,

baculovirus and retrovirus) Shuttle and expression vectors. properties of expression vector for cloning in

prokaryotes. Formation of chimeric plasmids using restriction enzymes, linkers, adaptors and

homopolymer tailing. Construction of artificial vectors.

UNIT III Isolation, cloning and selection

Isolation of a desired gene. Selection of ideal vectors and host cells. inserts, vector insert ligation,

transfection, DNA library – genomic and cDNA. advantages & disadvantages, construction of genomic and

full length cDNA libraries. BAC libraries, Restriction mapping, RFLP, Methods for screening and selection of

recombinant clones and expression of cloned genes.

UNIT IV Molecular Markers and DNA Chip technology

General account, RAPD, RFLP, AFLP, Simple Sequence Repeates (SSRs) or Microsatellites. DNA microarrays.

UNIT V Techniques

Principles and applications of blotting techniques : Southern, Northern, western, dot blot Polymerase

Chain Reaction – principle and application, RT-PCR, anchor PCR, inverse PCR, Asymmetric PCR.

oligonucleotide synthesis, site directed mutagenesis, insitu hybridization DNA fingerprinting and

footprinting. Chromosome walking and jumping. Antisense RNA technology.

UNIT VI Sequencing and synthesis of genes

List of suggested reading: 1. Recombinant DNA principles and Methodologies – James J. Greene

2. Molecular Biotechnology – Glick and Pasternak

3. Principles of Gene Manipulation – R. W. Old and S. B. Primrose

4. Genetic Engineering Fundamentals – Kammermeyer and Clark

5. Recombinant DNA. Watson et al.

6. Gene cloning: An introduction. T A Brown

7. Molecular cloning: A Laboratory Manual. Sambrook and Russell

8. Molecular Biology and Genetic Engineering. Gupta, P.K.

6. Molecular Biotechnology. Glick and Pasternak.



7. Biotechnology. Singh, B. D.



7003 (BT-303) ANIMAL BIOTECHNOLOGY

UNIT I Animal Cell Culture (Equipments, Consumables & Media)

Equipments and materials used in animal cell culture. Natural media, Synthetic media. Role of serum in cell

culture. Advantages and disadvantages of serum in culture medium. Advantages of serum free and law

serum media. Growth factors. Growth curve. Procedures for coating of culture surface. Scaling up of animal

cell cultures. Cytotoxicity and viability assays. Measuring parameters of growth. Dissagregation of tissue

and primary culture. Maintenance of cell culture and cell separation. Valuable products from cell culture.

UNIT II Sterilization and Lab Safety

Traditional Methods of Laboratory Safety & Sterilization. Special Methods used in Animal Biotechnology

Laboratory & Laboratory Layout.

Unit-III Cell Culture Technology

Biology and characterization of the cultured cells, measuring parameters of growth. Basic techniques of

mammalian cell culture in vitro; desegregation of tissue and primary culture; maintenance of cell culture;

cell separation & Tissue disaggregation. Scaling-up of animal cell culture Cell synchronization. Cell cloning

and micromanipulation. Cell transformation. Application of animal cell culture. Cultivation of Viruses

Unit-IV Stem Cells

Basics, embryonic & adult stem cells, Transdifferentiation, Applications, Ethical issues. Cell culture based

vaccines. Organ and histotypic cultures. Three dimensional culture and tissue engineering.

Unit-V Transgenic Animal Unit-I

Transgenic animal and their applications. Transgenic mice, sheep, goat, Rabbits, Cattle, Pigs and fish.

Animal Bioreactors and molecular pharming. Animal cloning. Ethical and social issues related to transgenic

animals.

Unit-VI Transgenic Animals-II

In vitro fertilization, ET and cloning. Transgenic Animals and Transfection Methods Introduction,

Objectives of gene transfer. Vectors, gene constructs. Promoters and reporter or marker genes. Transfection

Methods, Gene transfer through microinjection, Retroviral infection, Targeted gene transfer. Gene

disruption, gene replacement. Transgene integration. Gene therapy.

UNIT VII Applications of Animal Biotechnology

Biotechnology in Pest control, Aquaculture and sericulture. Role of biotechnology in biodiversity

conservation. Examples of Some Important Transgenic Animals.Application of molecular markers for

improvement of live stock, Detection of transgenics, Construction of Molecular Maps. Prospects of

transgenic livestocks. Embryo and gamete sex selection; nuclear transplantation, sperm separation,

embryonic sex selection.


1. Gene VII. Lewin, B.


BUNDELKHAND UNIVERSITY, JHANSI 2. Culture of Animal cells. Freshney, R.T. (2000) Alan R. Lirs Inc. New York.

3. Biotechnology and genomics. (2008) Gupta, P.K. Rastogi Publications Meerut.

4. Molecular Biology and Genetic engineering. (2008) Gupta, P.K.

5. Text book of Biotechnology. Das, H.K.

6. Biotechnology. Singh, B. D. (2008) Kalyani Publishers, New Delhi.

7. Fundamentals of Genetics. 2004. Singh. B. D. Kalyani Publishers. New Delhi.

8. Bioethics and Biosafety in Biotechnology. Sreeramulu, V. (2007) New Age International Publishers. New Delhi.



7004 (BT-304) PLANT BIOTECHNOLOGY

UNIT I Micropropagation

Historical background, Basic techniques of plant cell tissue culture, Surface sterilization, Aseptic tissue transfer,

Concept of totipotency.

Nutritional requirement of cell in vitro- Various type of nutrient media, Development of explants.

Methods of plant micro propagation. Ovary and ovule culture. Invitro pollination and fertilization. Embryo culture.

Embryogenesis and organogenesis and their practical applications.

UNIT II Haploid production

Haploids and their application. Anther and pollen culture. Monoploid production. Production of triploids through endosperm

culture. Somaclonal variation and applications. Single cell cultures and their application in selection of variants/mutants with or

without mutagen treatment.

UNIT III Protoplast and Embryo culture

Protoplast isolation and regeneration and its applications. Various methods of fusing protoplasts-Chemical, electrical.Somatic

hybridization. Selection system for hybrids. Role of protoplast culture and somatic hybridization in the improvement of crop

plants.

UNIT IV Crop Improvement through genetic engineering

Use of plant cell, protoplasts and tissue cuture for genetic manipulation of plants. Vectors for the constrcuction of transgenic

plants. Plasmid and plant virus vectors. Methods of gene transfer in to plant cells. Agrobacterium mediated. Direct gene

transfers. Cointegration and binary vectors. Gene gun. Microinjection. Integration and inheritance of transgenes.

Transformation of chloroplasts. Improved Crop productivity. Hybrid seed production. Biotic stress resistant plants. Engineering

Herbicide resistant plants. Direct strategy. Mutant psb A gene. Indirect strategy. Gene amplification and over expression.

Herbicide detoxifying genes. Bacterial detoxifying genes. Engineering viral diseases resistant plants. Coat protein expression

system. Insect resistant plants. Bt gene technology. Abiotic stress resistant plants. Plants resistant to drought, salt, heat, water

freezing etc. Improvement of nutritional quality. Seed storage proteins. Glycinin, conglycinin, legumin, phytohaemaglutinin,

phaseolin, prolamins, albumins and designer protein. Engineering for vitamins and iron deficiency. Transgenic plants suitable for

transport and shelf life. Terminator and traitor technology. Plant derived vaccines. Edible vaccines.

UNIT V Practical Concerns

Impact of genetically modified crops in genomic research and agriculture. Efficacy and Environmental concerns. Legislation for

transgenic plants.


1. Molecular Biology and genetic engineering. Gupta, P.K. (2008)

2. Biotechnology and Genomics. Gupta, P.K. (2008)

3. Molecular Biology in crop protection. Marshall, G and Walters, D.

4. Plant Biotechnology in Agriculture. Lindsey, K. and Jones, M .G.

5. Biosynthesis and Manipulation of Plant Products. Grierson, D.

6. Plants, genes, and Agriculture. Maarten J. Chrispeels and David E. Sadava

7. A text Book of Biotechnology R.C.Dubay

8. Biotechnology. Singh, B.D. (2008)

9. A text Book of Biotechnology. Kumar, H.D.



7005 (BT-305) LAB TECHNIQUES-III (A) MOLECULAR BIOLOGY

7006 (BT-305) LAB TECHNIQUES-III (B) INDUSTRIAL TECHNOLOGY

1. Microbial production of Ginger Beer

2. Production of spawn for white button mushroom

3. Use of alginate for cell immobilization

4. Isolation and Preservation of industrially important microorganisms

for microbial processes

7007 (BT-401) PROJECT/DISSERTATION

DEPARTMENT OF BIOTECHNOLOGY - Bundelkhand … · department of biotechnology bundelkhand...

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