DEPARTMENT OF BIOTECHNOLOGY JAWAHARLAL NEHRU ...

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DEPARTMENT OF BIOTECHNOLOGY

JAWAHARLAL NEHRU TECHNOL
OGICAL UNIVERSITY KAKINADA



M.Tech.

Biotechnology


T
he

Depa
rtment of Biotechnology at JNTU
Kakinada is dedicated to the advancement
of knowledge and is committed to quality education that combines
intense academic
study with the excitement of discovery. Excellent infrastructure and knowledge
resources provide the base. The department has initiated an M.Tech
.

programme in
Biotechnology. The programme equips its graduates to meet challenges in biotech

industries, while at the same time preparing them for research in frontier areas of
Biotechnology.


Eligibility criteria:

Candidates who have a four years Bachelor's degree

in
B.Tech
(Biotechnology)

,B.Pharmacy,

B.Tech(Chemical Engg), B.Tech(Biomedical Eng
g) or
Master's degree in
life
Science and
related subjects as one of the papers at the 10+2 level or
later

are eligible

to apply.




Jawaharlal Nehru Technological University, Kakinada

M.Tech (Biotechnology)

Course Structure

1 Semester

L

P

Total
Hours

Cr
edits

MBIO
-
101: Microbial Technology

4

-

4

4

MBIO
-
102: Metabolic Engineering

4

-

4

4

MBIO
-
103: Bioprocess Engineering

4

-

4

4

MBIO
-
104:Enzyme
Engineering& Fermentation
Technology

4

-

4

4

MBIO
-
105:Elective
-
I

Molecular fundame
ntals of Biology

Chemical plant &Equipment design

4




4

4

MBIO 106: Elective
-
II

Immunotechnology

Nano Biotechnology

4




4

4

MBIO
-
107: Micrbiology

&Immunology Lab


6

6

2

Total

2
4

6

3
0

26


III & IV Semesters

Industrial Training & seminar

Project Work


2
nd

Semester


L

P

Total
Hours

Credits

MBIO 201: Recombinant

D
NA Technology

4

-

4

4

MBIO
-
202: Bio separation Technology

4

-

4

4

MBIO
-
203: Bioinformatics, Genomics&
Proteomics

4

-

4

4

MBIO
-
204:

Bioreactor design


4

-

4

4

MBIO 205:Elective
-
II I

Stem cell Technology

Plant biotechnolog
y


4


4

4

MBIO
-
206: Elective
-
IV

Regulatory affairs and Clinical trials
/

Cancer biology

4


4

4

MBIO
-
207: r DNA technology & Bio
separation lab


6

6

2

Total

24

6

30

26

Code 101:

Microbial Technology

Unit

1
:
General Microbiology:
Principles,
Morphology & Cell structure of Prokaryotes and
Eukaryotes (Bacteria, fungi, algae and viruses, etc.); different culture techniques, isolation &
preservation methods, characteristics

of selecte
d group of microbes,
microbial nutrition and
growth, growth measurements techniques, energy & transduction in microbial systems,
transformation, conjugation
.

Unit

2:

Production of IFN, Interleukin,

HGF,

Vaccines
-
Production of various vaccines
,
plasmids as
vectors, Deletion, base pair substitutions and frame shift mutation, microbial
pathogenecity, Bioassays.

Unit 3:

Production of organic solvents

(
Beer, Wine), Production of fermented foods (
Cheese, Yoghurt), Production of organic acids (Citric acid, Acetic

acid and lactic acid),
Production of Amino acids (Glutamic acid and Lysine), Production of antibiotics (beta
-
lactams
-
Penicillins and Macrolids
-
erythromycin).




Unit4:

Improvement of industrial microorganisms by selection and mutation, importance of
genet
ic engineering in Microbial Biotechnology.

Unit 5
:

Major categories of industrial products (viz. alcohol, organic acids, enzymes,
antibiotics, vitamins, surfactants, polymers, microbial biomass, etc.)

Major biochemical
pathways (
Assimilatory, dissimilatory

& secondary pathway
s), Production technology (
Solid
and liquid phase fermentation, batch a
nd continuous fermentation
).

Unit 6
:

Basic concepts for design of a new protein/enzyme molecule, Design and
construction of novel proteins and enzymes, Visualisation

and inter
pretation of protein
structure.
Site directed mutagenesis

for specific protein function
-
Specific examples of
enzyme engineering.

Unit

7
:

Enzyme kinetics: Michaelis Menten Constant, Competitiy, Non
-
competitive,
uncompetitive inhibitions, Lineweave
r
-
Burke Plot

Stability of enzyme: Enzymes stabilization
by selection and genetic engineering, protein engineering.

Reaction environment building,
chemical modification,
intramolecular cross linking and immobilization.

Application of
biocatalysts: group tra
nsfers redox, elimination, isomerisation rearrangements, C
-
C bond
cleavage and formations. Application of enzymes in industry,

analytical purpose and medical
therapy.

Unit
8:

Bioleaching, Xenobiotic compounds, Bioremediation, Biostimulation,
Bioaugmentat
ion, Bioabsorption, and Phytoremediation.


TEXT BOOKS:

1. Microbial biotechnology: Glazer, A.N. and Nikaido, H. 1995 W.H. Freeman &


Company, New York.

2. A Textbook of Industrial Microbiology:
-

Wulf Creuger & Anneliese Creuger

3. Environmental Biotec
hnology by A.K. Chatterjee

4. Environmental Biotechnology: Basic concepts and applications by Indu Shekhar Thakur.

5. Environmental Biotechnology by S.N.Jogdand Himalaya Publishing


REFERENCES:

1. Foster C.F., John Ware D.A., Environmental Biotechnology,

Ellis Horwood Ltd.,


1987.

2. Karrely D., Chakrabarty K., Omen G.S., Biotechnology and Biodegradation,

3. Advances in Applied Biotechnology Series, Vol.4, Gulf Publications Co. London, 1989.

4. Bioremediation engineering; design and application 1995 J
ohn. T. Cookson, Jr.

5. Microbiology:
-

Prescott and Dunn. McGraw Hill, Inc.

6. Industrial Microbiology:
-

A. H. Patel.

7. Industrial Microbiology:
-

Casida.




Code 102: METABOLIC ENGINEERING


Unit

1
:

INTRODUCTION

Basic concepts of Metabolic Engineering
, Overview of cellular metabolism, Different models
for cellular reactions, induction , Jacob Monod model and its regulation, Differential
regulation by enzymes, Feed back regulation.

Unit

2
:

SYNTHESIS OF PRIMARY METABOLITES

Amino acid synthesis pathways

and its regulation at enzyme level and whole cell level,
Alteration of feed back regulation, Limiting accumulation of end products.
Engineering for
L
-
Lysine Production by
Corynebacterium glutamicum
-
Metabolic Engineering of Pentose
Metabolism for Ethanol P
roduction


Unit

3
:

BIOSYNTHESIS OF SECONDARY METABOLITES

Regulation of secondary metabolite pathways, precursor effects, prophase, idiophase
relationship, Catabolite regulation by passing control of secondary metabolism, producers of
secondary metabolites
, applications of secondary metabolites.

Unit

4
:

BIOCONVERSIONS

Applications of Bioconversions, Factors affecting bioconversions, Specificity, Yields, Co
metabolism, Product inhibition, mixed or sequential bioconversions, Conversion of insoluble
substanc
es.

Unit

5
:

REGULATION OF ENZYME PRODUCTION


Strain selection, Genetic improvement of strains, Gene dosage, metabolic pathway
manipulations to improve fermentation, Feed back repression, Catabolite Repression,
optimization and control of metabolic activit
ies. The modification of existing
-

or the
introduction of entirely new
-

metabolic pathways

Unit

6
:

METABOLIC FLUX

Metabolic Flux Analysis:

Theory and applications
-

metabolic flux analysis of citric acid
fermentation,Experimental determination method
of flux distribution, optimization and
control of metabolic flux.

Integrating Methodologies of Molecular Breeding

and bioprocess
systems engineering.

Fundamentals of Metabolic control analysis:

Control coefficients and
the Summation Theorems,

Elasticity Co
efficients and the Connectivity Theorems,
Generalization of MCA Theorems

Unit

7
:

METABOLIC ENGINEERING WITH BIOINFORMATICS

Metabolic pathway modeling, Analysis of metabolic control and the structure metabolic
networks, Metabolic pathway synthesis algorith
ms,

Unit

8
:

APPLICATIONS OF METABOLIC ENGINERRING

Application in pharmaceuticals, chemical bioprocess, food technology, agriculture,
environmental bioremediation and biomass conversion.

TEXT BOOKS:

1. Wang.D.I.C Cooney C.L., Demain A.L., Dunnil.P. Hump
hrey A.E. Lilly M.D.,
Fermentation and Enzyme Technology, John Wiley and sons 1980.

2. Stanbury P.F., and Whitaker A., Principles of Fermentation Technology, Pergamon Press,
1984.




















103
: Bioprocess Engineering


Unit 1:

Characteristic pro
perties of biological fluids, Non
-
linear properties of thermal death
of micro
-
organisms, Correlation of growth of Microbial cells and loss of biologically
oxidisable substances, different model
s of growth of micro
-
organisms

Unit 2:

Principles and mechanism
s of thermal stabilization by filtration, oxygen
-

supply and
demands in microbiological process, single and multiple bubbles aeration. On
-
ideality

and
RTD in Bioreactors

Unit3:

stability analysis; Analysis of multiple interacting microbial populations; Sta
bility of
recombinant cells; Physiology of immobilized cells; Packed
-
bed reactors; Fluidized
-
bed
bioreactors; Air
-
lift bioreactors; Bubble
-
column bioreactors; Immobilized
-
enzyme
bioreactors; Special react
ors for animal and plant cells

Unit 4:

On and off
-
li
ne sensors for a modern bioreactor,
integrated systems of bioreaction,
bioseparation biosensors
,
Characteristics of bio

products; Flocculation and conditioning of
broth;

Unit 5:

Mechanical separation; filtration, centrifugation and membrane based separatio
n; cell
disruption; protein precipitation and its separation; aqueous two
-
phase extraction;
Adsorption
-
desorption processes

Unit 6:

Chromatographic methods of separation based on size; charge hydrophobic
interactions, and metal ion affinity, etc.; Electrop
horesis; case studies.

Unit 7:

Chemical Engineering


Principles of biologists; Units and Dimensions, Introduction
to dimension analysis, Buckingham II Theorem, Unit operation and process in the Chemical
Industry, Fluid statics and Dynamics, Burnoulli’s e
quation,

Unit 8:


Newtonian and Non
-
Newtonian fluids, materials and energy. Balance on reactive
and non
-
reactive systems, principles of momentum, heat and mass transfer.

Texts

Books
:

1.


M. L Shuler and F. Kargi.,
Bioprocess Engineering,
Prentice Hall Inc
., 2002.

2.


P.M. Doran,
Bioprocess Engineering Principles
, Elsevier, 1995.

3.

P. B. Kaufman, L. J. Cseke, S. Warler, J. A. Duke, and H. L. Brielmann,
Natural Products
from Plants
, CRC Press



LLC, 1999.

References:

1.


H. J. Rehm


and G. Reed,


Bi
otechnology
-
A multi
-

Volume Comprehensive Treatise
, 2
nd

Ed, Vol 3, Wiley
-
VCH, 1993

2.


M. Moo
-
Young,
Comprehensive Biotechnology,
Vol. 2, Pergamon Press, 2004.

3.


F. Dicosmo and M. Missawa,
Plant Cell Culture Secondary Metabolism: Towards
Industrial A
pplication
. CRC LLC, 1996.


























Code

104
: EnzymeEngineering
& Fermentation Technology

Unit
-
1:

Kinetics and mechanism of enzyme action:

Classification of enzymes: quantification
of enzyme activity and specific activity. Estimation of Mech
aelis Menten parameters, Effect
of pH and temperature on enzyme activity, kinetics of inhibition. Modeling of rate equations
for single and multiple substrate reactions.

Unit
-
2:

Immobilized

enzyme reactions:

Techniques of enzyme immobilization
-
matrix
entra
pment, ionic and cross linking, column packing; Analysis of mass transfer effects of
kinetics of immobilized enzyme reactions; Analysis of Film and Pore Diffusion Effects on
Kinetics of immobilized enzyme reactions; calculation of Effectiveness Factors of
immobilized enzyme systems; Bioconversion studies with immobilized enzyme packed
-
bed
reactors.

Unit
-
3:

Mass Transfer Effects in Immobilized Systems:

Analysis of Film and Pore diffusion
effects on kinetics of immobilized enzyme reactions; Formulation of dim
ensionless groups
and calculation of effectiveness factors.

Unit
-
4:

Application of Enzymes:

Extraction of commercially important enzymes from
natural sources;
commercial

applications of enzymes in food, pharmaceutical and other
industries; enzymes for diag
nostic applications. Industrial production of enzymes. Use of
enzymes in analysis
-
types of sensing
-
gadgetry and methods. Case studies on application


chiral conversion, esterification etc.,

Unit
-
5:

Enzyme Biosensors:

Applications of enzymes in analysis; D
esign of enzyme
electrodes and case studies on their application as biosensors in industry, healthcare and
environment.

Unit
-
6:

Media design for fermentation processes:

Medium requirements for fermentation
processes, carbon, nitrogen, minerals, vitamins an
d other complex nutrients, oxygen
requirements, medium formulation of optimal gr
owth and product formation
. Medium for
plant cell culture and animal cell culture. Medium design of commercial media for industrial
fermentations
-
Plackett burman design, respon
se surface methodology, simplex design,
continuous cultivation method to determine the kinetic parameters of medium optimization.
Case studies on each medium design methods.

Unit
-
7:

Kinetics of microbial and product formation:

Phases of cell growth in batc
h cultures,
Fed batch and continuous cultures. Simple unstructured kinetic models for microbial growth,
Monod model, Growth of filamentous organisms & yeast. Growth associated (primary) and
non
-
growth associated (secondary) product formation kinetics, Leud
eking
-
Piret models,
substrate and product inhibition on cell growth and product formation.

Unit
-
8:

Fermentation technology:

Case studies on production of Lactic acid, Glutamic acid,
Pencillin, Microbial Lipase and Protease, Recombinant insulin. Case studie
s should deal with
strain improvement, medium designs, process optimization etc.

REFERENCES:

1.

Blanch, H.W., Clark, D.S. Biochemical Engineering, Marcel Dekker, 1997.

2.

Lee, James M. Biochemical Engineering, PHI, USA.

3.

Bailey J.E. & Ollis, D.F. Biochemical Engi
neering Fundamentals, 2
nd

Ed., McGraw
Hill, 1986.

4.

Wiseman, Alan. Hand Book of Enzyme Biotechnology, 3
rd

ed., Ellis Harwood 1995.

References:

1.

Bailey J.E. & Ollis, D.F. Biochemical Engineering Fundamentals, 2
nd

Ed., McGraw
Hill, 1986.

2.

Shuler, M.L. and Kargi,

F. Bioprocess Engineering: Basic concepts, 2
nd

ed. Prentice
-
Hall, 2002.

3.

Doran Pauline M, Bioprocess Engineering Principles, Academic Press, 1995.

4.

Stanbury, P.F., Stephen J. Hall & A. Whitaker, Principles of Fermentation
Technology, Science and Technology
Books.











Code 105:

Elective
-
I
Molecular fundamentals of Biology



Unit 1
. Molecular basis of life


an introduction. Experimental Pro of DNA and RNA as
genetic
,
Material; Ncleic Acids Structure and functions of DNA and RNA Wats on and Crick
model o
f DNA and other forms for DNA (A and Z) Functions of DNA and RNA including
ribozymes .


Unit 2
. DNA Replication: Prolaryotic and Eukaryotic


Enzymes and proteins involved in
replication, Theta miodel and Rolling circle model.


Unit 3
. DNA Repair: Causes
and mechanism
-
photoreactivation, excision repair, mismatch
repair, SOS repair; Recombination in prokaryotes Transformation, Conjunction and
Transduction.


Unit 4
. Structure of Prokaryotic and Eukaryotic gene
-

genetic code, Properties and Wobble

Hypothesis;

Transcription in prokaryotes and Eukaryotes Mechanism, Promoters and RNA
polymerase, transcription factors, Post transcriptional modifications of eukaryotic mRNA.


Unit 5
. Translation: Mechanism of translation in Prokaryotes and Eukaryotes, Post
transla
tional modifications of proteins.


Unit 6
. Regulation of Gene expression, Regulation of Gene expression in Prokaryotes


Operon concept (Lac and Tryp) Regulation of Gene expression in Eukaryotes
-

transcriptional activation, galactose metabolism in yeast.



Unit 7
. Gene organization and expression in Mitochondria and Choloroplasts.


Unit 8
. Insertional elements and transposons. Transposable elements

in Maize and
Drosophila.


Books:

1. Molecular Biology, Turner, P.C., McLennan, A.D. Bates & White, M.R.H., 2
nd Ed. Viva
Books.

2. Elliot, Biochemistry & Molecular Biology, OUP

3. Watson, Molecular Biology of Gene, Pearson Education

4. Cell and Molecular Biology, Rastogi, S.C., New Age International.























Code 105: Elective
-
I Chemical plant &Equi
pment design

U
nit
-
1 :
Introduction; development of flow diagrams from process description, material and
energy balance, sizing of equipment, design preliminaries, design codes, MOC selection
procedure, fabrication methods and testing methods.


U
nit

-
2:
Str
esses in thin and thick walled shells, theories of failure, design of storage vessels.

Unit
-
3:
Design of pressure vessels

Unit
-
4:
Design of shell and tube heat exchangers

Unit
-
5:
Design of single effect evaporator.

Unit
-
6:
Design of distillation and absorp
tion columns.

Unit
-
7:
Design of batch reactor, CSTR and PFR

Unit
-
8:
Optimum pipe diameter.

TEXT BOOK:

1. Chemical Engineering: Vol.6, Coulson J.M. and Richardson J.F., Pergamon Press 1983


REFERENCES:

1. Process Equipment Design, M.V. Joshi

2. Process Equi
pment Design
-
Vessel Design: Brownell L.E., Wiley Eastern Ltd.,(1986)

3. Introduction to Chemical Equipment Design
-
Mechanical Aspects: Bhattacharya B.C., CBS
Publishers, 1991

4. Process Heat Transfer: Kern Q., McGraw Hill book Co. Inc.,1982

5. Mass Transfer

Operations

:

Treybal R.E., MGH Book Co.Inc, 1982

6. Chemical Engineering Hand Book, Perry, 5th Ed.




Code 106: Immunotechnology

Unit 1
:
Immunology
-

fundamental concepts and anatomy of the immune system

Components of innate and acquired immunity; Phagocyt
osis; Complement and Inflammatory
responses;

Haematopoesis; Organs and cells of the immune system
-

primary and secondary
lymphoid organs; Lymphatic

system; Lymphocyte circulation; Lymphocyte homing; Mucosal
and Cutaneous associated Lymphoid

tissue.(MALT&CA
LT);


Unit 2:

Mucosal Immunity; Antigens
-

immunogens, haptens; Major Histocompatibility

Complex
-

MHC genes, MHC and immune responsiveness and disease susceptibility, HLA
typing
,

Immunoglobulins
-
basic structure, classes and subclasses of immunoglobulins
,
antigenic determinants;

Multigene organization of immunoglobulin genes; B
-
cell receptor;
Immunoglobulin superfamily; Principles

of cell signaling.


Unit 3
:
Immunological basis of self

non
-
self discrimination; Kinetics of immune response,
memory;B cell m
aturation, activation and differentiation; Generation of antibody diversity;
T
-
cell maturation, activation

and differentiation and T
-
cell receptors; Functional T Cell
Subsets; Cell
-
mediated immune responses, ADCC.


Unit 4
:
Cytokines
-
properties, receptors a
nd therapeutic uses; Antigen processing and
presentation
-

endogenous
,
antigens, exogenous antigens, non
-
peptide bacterial antigens and
super
-
antigens; Cell
-
cell co
-
peration,Hapten
-
carrier system
.


Unit 5 :
Antigen
-
antibody interactions

Precipitation, agglut
ination and complement
mediated immune reactions; Advanced immunological techniques

-

RIA, ELISA, Western
blotting, ELISPOT assay, immunofluorescence, flow cytometry and immunoelectron

microscopy; Surface plasmon resonance, Biosenor assays for assessing li
gand

receptor
interaction, CMI

techniq
ues
-

lymphoproliferation assay.


Unit 6
:
Mixed lymphocyte reaction, Cell Cytotoxicity assays, Apoptosis,Microarrays,
Transgenic mice, Gene knock outs
,
Active and passive immunization; Live, killed, attenuated,
sub unit

vaccin
es; Vaccine technology
-

Role.


Unit 7
:
properties of adjuvants, recombinant DNA and protein based vaccines, plant
-
based
vaccines, reversevaccinology; Peptide vaccines, conjugate vaccines; Antibody genes and
antibody engineering
-

chimeric andhybrid m
onoclonal antibodies; Catalytic antibodies and
generation of immunoglobulin gene libraries.


Unit 8
Clinical Immunology
:
Immunity to Infection : Bacteria, viral, fungal and parasitic
infections (with examples from each group);

Hypersensitivity


Type I
-
IV;

Autoimmunity;
Types of autoimmune diseases; Mechanism and role of CD4+ T

cells; MHC and TCR in
autoimmunity; Treatment of autoimmune diseases; Transplantation


Immunological

basis of
graft rejection; Clinical transplantation and immunosuppressive therapy
; Tumor immunology


Tumor antigens; Immune response to tumors and tumor evasion of the immune system,
Cancer

immunotherapy; Immunodeficiency
-
Primary immunodeficiencies, Acquired or
secondary immunodeficiencies.



TEXT BOOKS:

1.

E. Roitt Essential Immunology,

Vaccines conventional, Subnit and recombinant,
antidiotypic vaccine, Blackwell Scientific publications, Oxfold, 1991.

2.

K
uby Immunology, 5
th

Edition. Richard A Goldsby, Thomas J kindt Barbara A
Osborn . W H Freeman and Company.

3.


Kuby, RA Goldsby, Thomas J.
Kindt, Barbara, A. Osborne Immunology, 6th Edition,
Freeman, 2002.

4.

2. Brostoff J, Seaddin JK, Male D, Roitt IM., Clinical Immunology, 6th Edition,
Gower Medical Publishing, 2002.

5.

3. Janeway et al., Immunobiology, 4th Edition, Current Biology publications.,

1999.

6.

4. Paul, Fundamental of Immunology, 4th edition, Lippencott Raven, 1999.

REFERENCES:

1.

Benjamin E and Leskowitz S, immunology A short Course. Wiley LISS NY,
1991. ELISA Immunological Technological Techniques. DNA vaccines
Immunotechnology

2.

2. Cellular
Molecular Immunology. Abul Abbas and Lichman. 2003.



Code 106:

Nano Biotechnology



Unit 1:

Introduction to Nano Biotechnology
:
Background and Definition of Nano
biotechnology
-
Significance.

Supramolecular Chemistry: Definition and examples of the main
in
termolecular forces used

in supramolecular chemistry. Self
-
assembly processes in organic
systems. Main

supramolecular structures.


Unit 2:
Chemical approaches to nanostructured

materials
-
Molecular Building Blocks to
Nanostructures. Nanoscaled Biomolecules
-
Nucleic Acids and Proteins. Chemical Synthesis
of Artificial Nanostructures. Structural Control to Designed Properties and Functions.


Unit 3:
Nano materials
:
introduction
-
preparation
-
chemical vapor deposition
-
sol gels
-
electrodeposition
-
ballmilling

uses
and applications
.


Unit 4:
Nanofabrication:

Introduction
-
Basic techniques
-
MEMS fabrication techniques
-
nanofabrication techniques
-
Equipment and processes needed to fabricate nano devices and
structures such as bio
-
chips


Unit 5:
Bottom
-
up (building from mo
lecular le
vel) and top
-
down (breakdown of
microcrystalline

materials) approaches.



Unit 6:
Biologically
-
Inspired nanotechnology basic biological concepts and principles that
may lead

to the development of technologies for nano engineering systems. Coverag
e will be
given to

how life has evolved sophisticatedly.


Unit 7:
M
olecular nanoscale engineered devices, and discuss

how these nanoscale
biotechnologies are far more elaborate in their functions than most

products made by humans.



Unit 8:
Instrumentation

for nanoscale characterization. Instrumentation required for
characterization of properties on the nanometer scale. The measurable properties and
resolution limits of each technique, with an emphasis on measurements in the nanometer
range.




TEXT BOOKS:

1. Supramolecular Chemistry by Jean
-
Marie Lehn, Wiley VCH, 1995

2. Supramolecular Chemistry by Jonathan Steed & Jerry Atwood, John Wiley & Sons, 2004

3. Intermolecular and Surface Forces by Jacob Israelachvil, Academic Press, London, 1992i.













































Code 107:Lab on Microbiology&Immunology


1. Selection of animals, Preparation of antigens, Immunization and methods of bleeding,
Serum

separation, Storage.

2. Antibody titre by ELISA method.

3. Double diffusion, Immuno
-
electro
phoresis and Radial Immuno diffusion.

4. Complement fixation test.

5. Isolation and purification of IgG from serum or IgY from chicken egg.

6. SDS
-
PAGE, Immunoblotting, Dot blot assays

7. Blood smear identification of leucocytes by Giemsa stain

8. Separati
on of leucocytes by dextran method

9. Demonstration of Phagocytosis of latex beads

10. Separation of mononuclear cells by Ficoll
-
Hypaque

11. Flowcytometry, identification of T cells and their subsets

12. Lymphoproliferation by mitogen / antigen induced

13.

Lymphnode Immunohistochemistry (direct and indirect peroxidase assay)

14. Hybridoma technology and monoclonal antibody production.

15. Immunodiagnostics using commercial kits
















Code 201: Recombinant DNA Technology

Unit 1
:
DNA Structure and p
roperties; Restriction Enzymes; DNA ligase, Klenow enzyme,
T4 DNA polymerase,

Polynucleotide kinase, Alkaline phosphatase; Cohesive and blunt end
ligation; Linkers; Adaptors;

Homopolymeric tailing; Labeling of DNA: Nick translation,
Random priming, Radioac
tive and non
-
radioactive
.


Unit 2:
probes, Hybridization techniques: Northern, Southern and Colony hybridization,
Fluorescence in situ
,
hybridization; Chromatin Immunoprecipitation; DNA
-
Protein
Interactions
-
Electromobility shift assay;DNaseI footprinting; M
ethyl interference assay
.


Unit 3:
Cloning Vectors

Plasmids; Bacteriophages; M13 mp vectors; PUC19 and Bluescript
vectors, Phagemids; Lambda vectors;

Insertion and Replacement vectors; Cosmids; Artificial
chromosome vectors (YACs; BACs); Animal Virus

deriv
ed vectors
-
SV
-
40; vaccinia/bacculo
& retroviral vectors;

Expression vectors; pMal; GST.


Unit 4:
Omitted

vectors; Protein purification; His
-
tag; GST
-
tag; MBP
-
tag etc.; Intein
-
based
vectors; Inclusion bodies;Methodologies to reduce formation of inclusion bo
dies; aculovirus
and pichia vectors system, Plant basedvectors, Ti and Ri as vectors, Yeast vectors, Shuttle
vectors
.


Unit 5:
Cloning Methodologies
:
Insertion of Foreign DNA into Host Cells; Transformation;
Construction of libraries; Isolation of mRNA

and

total RNA; cDNA and genomic libraries;
cDNA and genomic cloning; Expression cloning; Jumping and

hopping libraries;
Southwestern and Far
-
western cloning; Protein
-
protein interactive cloning and Yeast

two
hybrid system; Phage display; Principles in maximiz
ing gene expression
.


Unit 6:
PCR and Its Applications
:
Primer design; Fidelity of thermostable enzymes; DNA
polymerases; Types of PCR


multiplex, nested, reverse

transcriptase, real time PCR,
touchdown PCR, hot start PCR, colony PCR, cloning of PCR produc
ts; Tvectors;

Proof
reading enzymes; PCR in gene recombination; Deletion; addition; Overlap extension; and

SOEing; Site specific mutagenesis; PCR in molecular diagnostics; Viral and bacterial
detection; PCR based
,
mutagenesis, Mutation detection: SSCP, DGGE
, RFLP, Oligo Ligation
Assay (OLA), MCC (Mismatch ChemicalCleavage, ASA (Allele
-
Specific Amplification),
PTT (Protein Truncation Test)
.


Unit 7:
Sequencing methods; Enzymatic DNA sequencing; Chemical sequencing of DNA;
Automated DNAsequencing; RNA sequenci
ng; Chemical Synthesis of oligonucleotides;
Introduction of DNA into mammalian
,
Genomic and c
-

DNA library, gene isolation, gene
cloning, expression of cloned gene DNA labeling by radioactive and non
-
radioactive
methods. DNA sequencing

chemica
l cleavage an
d dideoxy methods


Unit 8:
DNA finger printing, and their applications. Site
-
specific and oligonucleotide
directed mutagenesis, antisense and ribozyme technology; genetic diagnosis, gene transfer
technologies, transgenics; gene therapy.


Books:

1.

B. R. Gl
ick and J. J. Pasternak,
Molecular Biotechnology: Principles and Applications of
Recombinant DNA
, ASMPress : 3
rd
Edition,2003

2. S. Primrose, R. Twyman, B. Old, and G. Bertola,
Principles of Gene Manipulation and
Genomics
, Blackwel PublishingLimited;7
th
Edit
ion,2006

References:

1.

B. Alberts, A. Johnson, J. Lewis, M. Raff, K and R. P. Walter,
Molecular Biology of the
Cell
,

4
th


Edition,

Garland,

2002

2.


J. Hammond, P. McGarvey and V. Yusibov,
Plant Biotechnology: New Products and
Applications
,

Sprin
ger,1999.








Code202: Bio
-
separation Technology


Unit 1
:
Basic concepts of Bio
-
separation Technology, Separation characteristics of proteins
and enzymes


size, stability,

properties; purification methodologies Characteristics of bio
-
products; Floccul
a
tion and conditioning of broth.


Unit 2:
O
verview of reaction processes involved in separation, numerical examples
illustrating the process. Mechanical separation processes; Filtration at constan
t pressure and
at constant rate.


Unit 3:
E
mpirical equations

for batch and continuous filtration, centrifugal and

cross
-
flow filtration, Centrifugation: basic principles, design characteristics; ultracentrifuges::
principles and applications.


Unit 4:
Techniques Involved in Separation Processes

Foam
-
fractionation;
Solvent extraction
of bio
-
processes, aqueous two
-
phase extraction, adsorption
-
desorption process;

Salt
precipitation; Chromatographic separation based on size,


Unit 5:
C
harge hydrophobic interactions and metal ion affinity.Affinity chromatography,
inhibi
tors: their preparation and uses, method of linkages, Electrophoresis SDS
-
PAGE

(Polyacrylamide Gel), horizontal and vertical type, methods, case studies.


Unit 6:
Membrane based separation processes

Micro
-
filtration, Reverse osmosis,
Ultrafiltration and af
finity ultrafiltration, concentration polarization, rejection, flux
.


Unit 7:
E
xpression, membrane modules, dead
-
ended and cross
-
flow mode, material balances
and numerical problems.


Unit 8:
Industrial Applications
:
Industrial aspects of separation of bio
-
molecules, Material
balances, mathematical analysis and modelling: Case studies.


References: I

1. Schuler & Kargi, Bio
-
process Engg. PHI

2. Keith Wilson and John Walker, Practical Biochemistry

Principles and Techniques,
Cambridge, 5
th

Ed.2000

3. Coulson &

Richardson’s Chemical Engineering


Volume 3 (Chemical and Biochemical
Reactors and

process controls) ed. Richardson, J.F., Peacock, D.G., First Indian ed. Asian
Books Pvt. Ltd. 1998.

4. Bailey & oils, Biochemical Engg. Fundamentals, McGraw
-
Hill, 1990

5.
Geankoplis, C.J. Transport Processes and Unit Operations Prentice Hall of (I) 3rd ed. 1997.

6. Mukhopadhyay, S.N. Process Biotechnology Fundamentals, Viva Books Pvt. Ltd. 2001.

7. Muni & Cheryan, Handbook of Ultrafiltration
.


References: II

1.Perry, Chilto
n & Green, Chemical Engineers’ Handbook, McGraw
-
Hill

2. Ho, W.S.W. and K. K. Sirkar, Membrane Handbook, Van Nostrand Reinhold,

N.Y. (1992)
.
































Code 203: Bioinformatics, Genomics& Proteomics

Unit 1:
Introduction to Genomic data a
nd Data Organization: Sequence Data Banks


Introduction to sequence date banks

protein sequence data bank. NBRF
-
PIR, SWISSPROT,
Signal peptide data bank, Nucleic acid sequence data bank

GenBank, EMBL nucleotide
sequence data bank, AIDS virus sequence da
ta bank.


Unit 2:
Applications in Biotechnology: Protein classifications, Fold libraries, Protein
structure prediction: Fold recognition (threading), Protein structure predictions : Comparative
modeling (Homology), Advanced topics: Protein folding, Protei
nligand interactions,
Molecular Modeling & Dynamics, Drug Designing.


Unit 3:
Secondary Structure predictions; prediction algorithms; Chao
-
Fasman algorithm,
Hidden
-
Markov model, Neural Networking. Tertiary Structure predictions; prediction
algorithsms; Cha
o
-
Fasman algorithm, Hidden
-
Markov model, Neural Neworking.


Unit 4:
Introduction to protemics and protein engineering
-

Protein prefractionation and
sample preparation
-

Two dimensional electrophoresis (2
-
D PAGE)

-

Protein identification

-

Post translation
al modification


Unit 5:
Functional

Genomics

and analysis of gene expression

-

Proteomics and drug delivery

-

Reverse genetics

Comparing transcriptomes
-

subtractive hybridization, differential display, SAGE,
Microarrays Genetic diseases in humans, Human G
enome project, Genetic counseling,
Genetics and society.


-

Transcription and replication of negative strand viruses


Unit 6:
Protein engineering and transfer RNA world
; Proteome analysis
-

Essential
requirements for protein synthesis
,
Role of messenger RNA
,
SNIJRPS and Introns
,
Translation
, Proteome analysis:
The impact of stable isotope labeling: Sample preparation, 2
-
D gel separation and analysis, Mass spectrometry: pr
otein identification using MS

data, Gel
matching, Protein chips and applications.



Unit

7:
Phylogeny
:
Concepts of systematic, Molecular evolution, Definition and Different
types of phylogenetic trees, Dendrograms and interpretations, phylogenetic analysis


Unit 8:
Genomics in Biopharmaceutical Industry Functional Genomes
-
Pharmacogentics
-
Geno
mics in relation to molecular Diagonosis
-
Molecular Therampeutic technologies.

Drug
Discovery
-
Overview, Stages of drug discovery, Functional genomics tools, Functional
Proteomics tools, Bioinformatics applications in experimental biotechnology.



References
:

1. Lesk, Introduction to Bio Informatics, OUP

2. Developing Bioinformatics Computer Skills, Cynthia Gibas and Per Jambeck, 2001 SPD

3. Introduction to Bioinformatics, Atwood, Pearson Education

4. Beginning Perl for Bio
-
informatics, Tisdall, SPD

5) H.D. K
umar, Molecular Biology,2nd edition, Vikas Publishing House pvt ltd

6) B.Alberts,D.Bray, J.Lewis et al, Molecular Biology of the Cell, Garland Pub. N.Y 1983

7) Concepts in Biotechnology D. Balasubramaniam, Bryce, Dharmalingam, Green,
Jayaraman

Univ. Press,

1996

8)
S.Sahai, Genomics and Proteomics, " Functional an Computational Aspects ",

Pienum Publications, 1999.











Code
204: Bioreactor design

Unit 1
:
Introduction: Bioreactor function, utility, types of Bioreactors. Modes of Bioreactor
operations,

Main components of the Bioreactor and their function. Introduction Methods of
Aeration, Surface Aeration, shake flasks, Mechanical stirred Bioreactors, Enzyme catalysis in
CSTR. Cell death in batch reactor, endogenous metabolism, maintenance, product and
substrate inhibition on chemostat.


Unit 2:
Bioreactors and design features; Batch reactor, chemostat CSTR, Plug flow Reactor,
Fed batch Reactor, Bubble column, Bubble generation at an orifice, bubble coalescence and
breakup, gas holdup, interfacial area,
immobile and mobile gas liquid interface, regimes of
bubbles, design of bubble columns, Cascade Reactor, air lift reactor, Fluidized bed
bioreactors, trickle bed reactors, immobilized bioreactors, recycle bioreactors.


Unit 3:
Gas
-
liquid mass transfer in c
ellular systems, basic mass transfer concepts, solubility
of gases (O2, CO2) in biological media, mass balances for two
-

phase Bioreactor. Mass
transfer
-

introduction to mass transfer between phases, mass transfer in porous solids,
quantifying mass transfe
r, mass transfer & experimental design. Oxygen transfer
-

introduction, oxygen transfer process, factor effecting kLa, interfacial area and oxygen
transfer, factors effecting the saturation concentration of oxygen, oxygen uptake.


Unit 4:
Determination of o
xygen transfer rates, mass transfer for freely rising or falling
bodies, forced convection mass transfer, overall kLa estimation and power requirements for
sparged and agitated vessels, mass transfer across free surfaces, other factors affecting kLa,
non N
ewtonian fluids, Heat transfer correlations, thermal death kinetics of microorganisms,
batch and continuous heat, sterilization of liquid media, filter sterilization of liquid media,
Air. Design of sterilization equipment batch and continuous.


Unit 5:
Mas
s transfer in agitated tanks, correlations with kLa in Newtonian and non
Newtonian liquid, power number. Experimental determination of kLa, static method,
Dynamic method, Chemical method and Electrochemical method. Power requirement for
mixing in aerated a
nd non aerated tanks, agitated and non agitated tanks for Newtonian and
Non Newtonian fluid. Mixing time in agitated reactor, residence time distribution, non ideal
reactor and multiphase bioreactor.


Unit 6:
Aeration and agitation in animal cell bioreacto
rs; Introduction, cell damage in animal
cell bioreactors, shear damage, bubble damage, methods of minimizing cell damage. Laminar
& Turbulent flow in stirred tank bioreactors, turbulent eddies, kolmogrov eddy size,
preventing vortex formation, off centre i
mpellers, Baffles. Control of bioreactor, strategy,
online and offline monitoring of bioreactors; computerized bioprocess control, scaling up and
scale down of mass transfer equipment and bioprocess, control of bioreactor, sensor used in
the bioreactor, pH
, O2,CO2 electrode. Online sensors for cell properties. Direct regulatory
control and cascade control mechanism.


Unit 7:
On
-
line data analysis for measurement of important physico
-
chemical and
biochemical parameters; Methods of on
-
line and off
-
line biomas
s estimation; microbial
calorimetry; Flow injection analysis for measurement of substrates, product and other
metabolites; State and parameter estimation techniques for biochemical processes. Case
studies on applications of FIA and Microbial calorimetry.


Unit 8:
Bioreactor strategies for maximizing product formation; Case studies on high cell
density cultivation and plasmid stabilization methods. Bioprocess design considerations for
plant and animal cell cultures. Analysis of multiple interacting microbial

populations


competition: survival of the fittest, predation and parasitism: Lotka Volterra model. Reactor
dynamics and stability. Reactors with non ideal mixing. Other types of reactors
-

fluidized bed
reactors, packed bed reactors, bubble column reactor
s, trickle bed reactors. Scale up by
geometry similitude, oxygen transfer, power correlations, mixing time.



TEXT BOOKS:

1) Bailey JE, Ollis DF; Biochemical Engineering fundamentals Year of Publication 1986.

2) Blanch HW and Clark DS: Biochemical Engineer
ing Marcel Decker Year of Publication
1987.

3) Introduction to Biochemical Engineering by DG Rao, Tata Mc Graw Hill, New Delhi.




REFFERENCE BOOKS:

1)

Wiseman, A: Handbook of Enzyme Biotechnology, 3
rd

Edition, Ellis Horwood
Publication, Year of Publication 1
999.

2)

Moser, A; Bioprocess technology, kinetics and reactors; Springer Verlag, Year of
Publication 1988.

3)

Schugerl K: Bellagart K H (Eds); Bioreaction Engineering, Modeling and control;
Springer
-

verlog, berlin year of Publication 2000.

4)

Biochemical Enginee
ring Principles and functions by Syed Trnveer Ahmed Inamdar,
PHI Learning Private limited.

5)

Moser,Anton, Bioprocess Technology: Kinetics and Reactors, Springer Verlag,1988.

6)

Bailey J.E. & Ollis,D.F. Biochemical Engineering Fundamentals,2
nd

ed., McGraw
hill,
1986

7)

Lee, James M. Biochemical Engineering, PHI, USA

8)

Atkinson, Handbook of Bioreactors Blanch, H.W. Clark, D.S. Biochemical
Engineering, Marcel Decker, 1999













Code 205
: Plant Biotechnology

Unit 1
:
Introduction of plant tissue culture and cell sus
pension culture, physico
-
chemical
conditions for propagation of plant cells

and tissues, composition of media, nutrient and
hormone requirement, continuous culture, techniques for immobilization of

plant cells,
continuous product recovery system using immo
bilized plant cell system.


Unit 2:
Plant tissue culture


product and recovery:

Primary and secondary metabolic products (phytochemicals) of plant cells, biosynthesis of
secondary metabolites of

Biotechnological importance, biotransformation for product
d
evelopment and selection of cell culture, process technology

with salient features for
specific products.


Unit 3:
Plant tissue culture


genetic engineering (a):

Structure and organisation of plant
genome, regulation of plant genome expression, transcript
ional, translational and post

transcriptional regulation of plant genome.

-

Transposons, chloroplast and mitochondrial
genome.


Unit 4:
Plant tissue culture


genetic engineering (b):

Transfer of nucleic acid to plant cells

-

Direct transformation by elect
roporation and particle gun bombardment. Agrobacterium, Ti
plasmid vector

Theory and techniques for the development of new genetic traits, conferring
resistance to herbicide, pesticide, plant

pathogens.


Unit 5:
Plant engineering towards development of enr
iched food products,

plant growth
regulators
, Molecular pharming.


Unit 6:
Introduction to primary and secondary metabolites, important pathway leading to the
biosynthesis of secondary metabolites (e.g. serpentine, shinkonin, diosgenin and cardenolids)
in
plants,

Unit 7:
Metabolic products produced by in vitro culturing of plant cells, selection of plant
cells/tissues for the production of a specific product, Culture system in secondary plant
product biosynthesis
-
batch continuous cultures and immobilized p
lant cells, iotransformation
of precursors by cell culturing.


Unit 8:
Extraction and analytical methods for the above four metabolites. Industries involved
in the production of plant secondary metabolites, Potential and future prospect of the
secondary m
etabolites production by plant cell culture techniques.



Text Books:

1.Cripeels, M.J. and Sadava, D.E., Plants, Genes and Biotechnology, jones and
Biotechnology, Jones and Bartlett Publishers (2
nd

Edition), 2003

2. Bhowjwani, S.S., Plant Tissue Culture: A
pplication and Limitations. Amsterdam,
Elsevier,1990.

3.Chartes, Cunningham and Andrew J.R. Porter, Recombinant Proteins from Olans
Production and Isolation of Clinically Useful Compounds (Methods in
Biotechnology),Humana Press,1997

4. Bemard R.Glick and J
ohn E. Thompson, Methods in Plant Molecular Biology and
Biotechnology, CRC Press, 1993.

5.I. Potrykus and G.Spangenberg, Gene Transfer to Plants (Spinger Lab Manual), Spinger
Verlag, 1997.

6. Peter m. Gresshoff, Plant Genome Analysis: Current Topics in Pla
nt Molecular Biology.
CRC Press, 1994.

7. John Hammond, Peter McGarvey, Vidadi Yusibov, Plant Biotechnology: New Products
and Applications, Springer, 1999.










Elective IV


CODE 206: REGULATORY AFFAIRS AND CLINICAL TRIALS


UNIT
-
1:
I
NTRODUCTION TO CLI
NICAL TRIALS:

a brief history of clinical trials
--

types of clinical trials
--

ethics of clinical trials
--

Clinical Trial Protocols


UNIT
-
2:
DESIGN AND ORGANISATION OF A CLINICAL TRIAL:

treatment
assignment methods: simple randomisation, blocked randomisat
ion, stratified randomisation
---
minimisation method
-

unequal randomisation
-

the size of a clinical trial
-
reporting results
-

Roles and responsibilities of clinical research personnel according to ICH
-
GCP


UNIT
-
3

:
MONITORING TRIAL PROGRESS:
INTRODUCTION
-

OUTCOME
MEASURES
-

c
ompliance, Dropouts
and
-
Intention
-
To
-
Treat
-

A
Taxonomy Of Dropouts
-

Interim Analyses In Clinical Trials
-

Group Sequential Procedures
---

A
Bayesian Perspective.

Audits In Clinical Research


UNIT
-
4:
BASIC ANALYSES OF CLINICAL TRIALS
: I
ntroduction
-

Brief Review
Statistics
-

Generalized Linear models: Models for Counts: Treat
ment of Familial
Adenomatous Pol
yposis (FAP) with a Non
-
Steroidal Anti
-
Inflammatory Drug
-
Ordinal
Response Models:A Clinical Trial in Lymphoma
-

Models with parallel
-

Non
-
parallel
models
-

Model estimation linear predictors
-

multiple endpoints
-

Analysis of Longitudinal
Data from Clinical Trials


UNIT
-
5:
HISTORY OF REGULATORY AFFAIRS
, Main concepts QSE
-

Regulatory
affairs for studies in human subjects
-

data needed
-

Li
censing authorities
-
roles and
responsibilities,ICH
-
GCP,FDA, EU Clinical Trial
-

Regulatory submissions and
Requirements for gaining approval of new products
-
Regulating control over marketing and
sales of medical products


UNIT
-
6

:
Documentation and Mainte
nance of records.


UNIT
-
7:
Drug & Cosmetics Act with special reference to schedule Y and M, schedule of
medical devices. Concept of total quality management, requirements of GMP, GLP, GCP,
Regulatory requirements of drugs and Pharmaceutical (USFD
-
NDA/ AN
DA)



UNIT
-
8:
INTELLECTUAL PROPERTY RIGHTS

patents, Trademarks, Copyrights,
Patents Act.


TEXT BOOKS:

1. Willing, S.W., & Stoker, Good Manufacturing Practices for Pharmaceuticals, Marcel
Dekker, New York.

2. Guarino, R.A., New Drug Approval Process, Marc
el Dekker, New York.

3. Drug & Cosmetic Act.

4. Patents Act.

5. Bansol, IPR Guidelines for Pharm students and Researchers.

6. Pisano
-
FDA Regulatory Affairs.

7. Phillip W. Grubb, Patents for Chemicals, Pharmaceuticals and Biotechnology



8.

Internation
al Clinical Trial, Volume 1 &2 Dominique

P.Brunier and Gerhardt Nahler,


Interpharm Press, Denver, Colorado

9
. Code of Federal Regulation by USFDA


Download


10. ICH
-
GCP Guidelines


Download

11
. Biosafety issues related to genetically modified orga
nism , Biotech Consortium India
Limited, New Delhi



12.
Brian S. Everitt.,

Statistical Aspects Of The Design And Analysis Of Clinical



Trials(Revised Edition)
-

World Scientific Publishing




















CODE 206: CANCER BIOL
OGY


Unit 1:

Introduction: Causes of cancer:
Physical, Chemical, Biological, Diet, Genetic &
Hereditary. Types of Cancer, Cell Cycle Events, Regulation of Cell Cycle, Signal Molecules,
Signal receptors, Mutations in signal molecules that alters cell cycle,

Tumour suppressor
genes, Role of tumour suppressor genes in control of cancer.


Unit 2:

Carcinogenesis:

Chemical Carcinogenesis: History, Examples of known to be human
carcinogens , Mechanism of carcinogenesis, Targets of Chemical carcinogens, DNA Adducts
,
Phase I & Phase II Metabolism of Chemical carcinogens, Physical carcinogens: X ray
radiation induced carcinogenesis, Asbestos induced carcinogenesis


Unit 3:

Molecular Biology Of Cancer
:

Detection of Oncogenes, Identification of Oncogenes,
Retroviral onc
ogenes, Growth factors and Growth factor receptors as oncogenes.


Unit 4:

Metastasis
: Significance of invasion: Heterogeneity of Metastasis, Metastatic
Cascade, Basement Membrane disruption, three step theory of invasion, Proteinases and
tumour cell invasi
on


Unit
5:

Cancer Screening & Detection
:
Screening Methods, Molecular detection of Cancer,
Low throughput assays, SSCP, CGH, Flowcytometry, Southern blot, RFLP, FISH, High
throughput techniques: sequencing, Microarrays. CT Scan, MRI Scan, PET Scan


Unit
6
:

Chemotherapy
: Mechanism of action of cytotoxic drugs, resistance to cytotoxic
drugs, Early and late side effects of chemotherapy, Cytotoxic drug discovery, Pre clinical,
clinical trials phase I, Phase II & Phase III, Anti Cancer agents
-

Alkylating agents
,
Antimetabolites, Topoisomerase inhibitors


Unit 7:

Radiation Therapy:

Radical Radiotherapy, Adjuvant radiotherapy, Palliative
radiotherapy, Gross Tumour Volume(GTV), Clinical Target Volume (CTV), Planned Target
Volume(PTV), Classical radiation biology,
Cellular Response to DNA Damage


Unit
8:

Immunotherapy
: Specific Immunotherapy: T Cells, Dendritic Cells, DC Vaccines,
Peptide vaccines, Non specific immunotherapy
-

cytokines


REFERENCES

Cell by Cooper

Cell and Molecular Biology by Lodish et al

Biology of
Cancer by Weinberg

Cellular and Molecular Biology of Cancer by Margaret Knowles and Peter Selby, Oxford
University Press








































Code 207:

r DNA technology & Bio separation lab


1. Isolation of genomic DNA from
Bacillus subti
lis*
genome.

2. PCR amplification of
scoC
gene and analysis by agarose gel electrophoresis

3. Preparation of plasmid, pET
-
28a from
E.coli
D
H5
and gel analysis.

4. Restriction digestion of vector (gel analysis) and insert with NcoI and XhoI

5. a. Vector and

Insert ligation

b. Transformation in E.coli DH5
.

6. Plasmid isolation and confirming recombinant by PCR and RE digestion.

7. Transformation of recombinant plasmid in
E.coli
BL21 (DE3) strain.

8. Induction of ScoC protein with IPTG and analysis on SDS
-
PAGE

9. Purification of protein on Ni
-
NTA column and analysis of purification by SDS
-
PAGE

10.

a. Random Primer labeling of scoC with Dig
-
11
-
dUTP

b. Southern hybridization of
B. subtilis
genome with probe and non
-
radioactive
detection.

*Any other bacterial str
ain can be used.