BMERF  INDIA

 

Biomedical Engineering Research Foundation  

RESEARCH INSTITUTE OF BIOTECHNOLOGY

AFFILIATED TO PERIYAR UNIVERSITY

 
 

 

1.  SHORT TERM CERTIFICATE COURSES

(TWO WEEKS / ONE MONTH)   

  1. ANIMAL TISSUE CULTURE TECHNIQUES

  2. BIOINSTRUMENTATION

  3. ELECTROPHORESIS & ELECTROBLOTTING

  4. FERMENTATION TECHNOLOGY

  5. GENE CLONING & EXPRESSION

  6. IMMUNOTECHNOLOGY

  7. PLANT TISSUE CULTURE

  8. PCR & GEL DOCUMENTATION

  9. BLOTTING & HYBRIDIZATION

  10. DNA / RNA PROBE DEVELOPMENT

Highlights of the  training Programs @ BMERF

  • receiving and consolidating samples without teaching kits,
  • DNA extraction with latest protocols,
  • PCR setup,
  • fragment resolution,
  • analysis of data,
  • communication of results to internal customers and
  • some troubleshooting.
  • the individual will also order supplies,
  • maintain the lab and automated equipments,
  • utilize an internal database to manage samples and
  • assist with the relocation of the lab.

About the instructors

              Every instructor in the program is a professional (B.Tech/M.Tech/Ph.D) now working in the biotechnology/biomedical industry — or   a college instructor teaching biotechnology / biomedical courses — or both.  One faculty is from France who is cross trained in both biotech and bioinformatics laboratories.

How to get to the classes?

        All classes are held at the Research Institute of Biotechnology conveniently located at Yercaud (hill station) foothills of  Salem, Haroor Main Road, Kuppanoor,  which is 10 miles from Salem City (South India).   However, molecular biological techniques are also taught at THE GENE TECH R&D UNIT, GOVT. INDUSTRIAL ESTATE, UDAYAPATTI, SALEM - 636 140. 

Should there be any query the candidate may also approach us easily in our ADMINISTRATIVE OFFICE, which is located in the THE GENE TECH BUILDING, GOVT. INDUSTRIAL ESTATE, UDAYAPATTI, SALEM - 636 140. 

         Please call +91 - 0427-2241422 / (0) 9443344299 / (0) 9442955315 if you have any query about the content of any of the courses described in this brochure.

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 ONE YEAR DIPLOMA program DETAILS

1.   DIPLOMA IN GENOME TECHNOLOGY -  2009-10

Paper I – General Paper

Unit I  : Life Skills & Communications:

01)       a. External and internal influences in one’s life.

02)       a . Meaning and process of coping

              b. Mechanism of coping

              c. Coping with physical change and sexuality

03)        a. coping with shyness, fear

             b. Coping with anger, criticism , failure

 04)       a. Self-Acceptance, Self-esteem

             b. Self-actualization

05)        a. Positive thinking- motivation

              b. Goal setting

06)        a. Problem solving

             b. Decision making

07)        a. Time Management

             b. Stress Management

08)        a. Leadership

             b. Team building

09)        a. Inter personal elements of communication

             b. Expression. Listening and responding

10)        a. Group Communication and Group discussion

 

 

Books for Reference

0    1)      “ Organisational Behaviour” by Jit S.Chandan

      2)      “Organisation Behaviours” by M.M.Varma and Adarval

      3)      “ How to be your own best friend” by Paul Mauck

 

  

Unit II  : Spoken English:  

 

Situational Skills: Greeting – Introducing oneself –Inviting, someone – Making requests Offering help –   Seeking permission –    Asking for advice – Expressing gratitude –Asking about remembering-   – persuading  - complimenting / Congratulating – Expressing sympathy – complaining – Apologizing – Making suggestions – Warning someone – Starting conversation with a stranger – Leaving someone for a short time – Ending a conversation – Asking for information – Asking for someone’s Opinion – Asking if someone’s sure –Asking someone to say something again checking that you have understood – Asking whether someone knows – Asking about  possibility –   Asking about preference – Asking if someone     is able to do something – Describing something – Some   useful expressions.   Building Vocabulary: One word substitution  - Synonyms - Homophones – Odd man out. Language Use: Identification of tenses and agreement of verbs. Statements, Negatives,  Interrogatives and Question tags. Rectification of errors in articles. Propositions and adverbs  – Comprehension – telegraphic Message.

 

 

Books for Reference:

 01) Spoken English for  you – Emerald Publishers – Authors:

           G.Radhakrishna Pillai ,K.Rajeevan.

 02)  Spoken English  - A Self Learing  Guide to Conversation Practice      

      Authors: V.Sasikumar, P.V.Dhamija

 

Unit III : Computer Usage and Applications:  

01) Introduction to Computer: Generation and History of Computers- Characteristics of Computer – Digital Computer - Analog - Computer – Hybrid Computer. Classification of Digital Computer: Micro Computer –Mini Computer – Mainframe computer .

02) Input Devices: Keyboard, Mouse, Light pen –Output Devices: Monitor, Classification of Monitors, Printers, Classification of printers –Study of    Secondary storage devices.

 03)     Hardware, Software, farmware, parts of CPU: ALU – Control Unit –Main   memory – RAM, ROM.

 04)     System Software: Interpreter, Assembler, Compilers –        Operating Systems: DOS – Booting System – Internal and   External Commands – Introduction to High-level languages.

 

05)Origin of Internet – Appanet – MODEM – ISP (VSNL) –  Upload, Download. E-mail: Origin of Worldwide web (WWW) –   Browsers – HTML     Basics.  

    Books for reference:

01)   Introduction to Computers by Radhakrishnan.  

02)    Fundamentals of Computers by V. Raja Raman.

 

Paper 2: Central dogma system

UNIT I

1.          Molecular nature and fine structure of gene         

Ψ     Central dogma of life

Ψ     The chemical nature of the gene:

Ψ     The structure of DNA

Ψ     The Watson-Crick proposal

2.       DNA replication:

Ψ     Semi conservative replication

Ψ     Replication in prokaryotes and eukaryotes

3.          Transcription:

Ψ     Basic processes

Ψ     Transcription in prokaryotes and eukaryotes

Ψ     Ribosomal RNA

Ψ     Transfer RNA and

Ψ     Messenger RNA

4.       Translation:

Ψ     Initiation

Ψ     Elongation and

Ψ   Termination

 UNIT II

Ψ     Gene as a unit of mutation and recombination

Ψ     Muton

Ψ     Recon

Ψ     Cistron

Ψ     Recombinant DNA expression systems and their applications

Ψ     Expression systems developed to produce recombinant proteins

Ψ     Insulin

Ψ     Hepatitis B vaccine

Ψ     Monoclonal antibodies

Ψ     New approaches to drug designing

UNIT III

Ψ     Origin of spontaneous mutation and it’s control

Ψ     Chemical mutagenesis by nitrous acid and hydroxylamine

 UNIT IV

Ψ     Mutagenesis by alkylating agents, intercalators

Ψ     Physical mutation by UV

UNIT V

1.     Point mutation

2.     Site directed mutagenesis:

Ψ    Cassette mutagenesis

Ψ    Oligonucleotide directed mutagenesis,

Ψ    PCR methods of site directed mutagenesis.

 

Paper 3: Recombinant DNA technology

UNIT I

1.     Cloning strategies

Ψ    Restriction endonucleases and its applications

Ψ    Isolation of DNA fragments

Ψ    Splicing the target DNA with the vector

Ψ    Strategies involved in introducing the cloned vector to host cell

Ψ    Selection or screening of recombinants

2.     Construction of gene libraries

Ψ    Genomic DNA libraries

Ψ    E. coli host for library construction in phage vectors

Ψ    Chromosomal walking

Ψ    Cutting DNA at very rare target site

Ψ    PFGE & FIGE

Ψ    Chromosomal jumping strategies

Ψ    Construction of genomic DNA libraries

Ψ    cDNA libraries and their applications

UNIT II

1.  Probe construction  - Definition

Ψ     Radioactive and Non-radioactive labeled probes

Ψ     Features of DNA probe

Ψ     Steps involved in probe construction

Ψ     Application

2.  Recombinants selection and screening

Ψ     Genetic methods – antibiotic resistance and nutritional markers

Ψ     Immunochemical methods - Western blotting / Immunoblotting

Ψ     Southern blotting, Northern blotting

Ψ     Nucleic acid hybridization

Ψ     Hybrid arrested translation

Ψ     Hybrid released translation

UNIT III

1.     Production of Proteins from cloned genes

2.     Translation in in-vitro Transcription and translation in in-vivo

3.     Lac Z promoter, l T 7 promoters and hybrids

4.     Dealing with fusion proteins

5.     Fate of the transcripts

6.     Efficiency of translation

7.     Fate of proteins after synthesis

8.     Gene cloning in medicinal and agricultural research

UNIT IV

1.  Intellectual property rights and patenting

Ψ     Constituents of patency

Ψ     Types of patency

Ψ     Patenting in different countries

Ψ     Patenting and fundamental research

Ψ     Patenting biotechnological inventions

 

UNIT V

1.  Biosafety containment facilities for genetic engineering experiments

Ψ     Physical containment

Ψ     Biological containment

Ψ     Risk evaluation

PAPER 4 : VECTOR construction and Gene transfer

UNIT I

1.  Extra chromosomal heredity

Ψ     Biology of plasmid

Ψ     Basic properties of plasmids

Ψ     Desirable properties of plasmids as cloning vehicles

Ψ     Usefulness of natural plasmids as cloning vehicles

UNIT II

1.  E. coli vectors

Ψ     Construction and characterization of a new cloning vehicle: pBR 322 (origin of plasmid pBR322)

Ψ     Examples of uses of plasmid   pBR322 as a vector

Ψ     Improved vectors derived from pBR322

UNIT III

1.  Cloning vectors for gram positive & gram negative bacteria

Ψ     Cloning in Bacillus subtilis

Ψ     Improved vectors for cloning in B. subtilis

Ψ     Cloning in Streptomyces

Ψ     E. coli vectors; plasmids

Ψ     Transformation

UNIT IV

1.  Transfer of genes by conjugation and transduction

Ψ     Sex factor F

Ψ     Generalized transduction

Ψ     Specialized transduction

UNIT V

1.  Phages:

Ψ     Bacteriophage l

Ψ     Essential features

Ψ     Promoters and control circuits

Ψ     Improved phage l vectors 

Ψ     Packaging phage l DNA in-vitro

Ψ     M13 phage

2.  Cosmid vectors

3.  Phasmid vectors

4.  Shuttle vectors

 

PAPER 5:  TRANSPOSABLE   ELEMENTS

 UNIT I 
Definition of transposons, Discovery,

 Early experiments of Mc. Clintock in maize,

 Transposon mutagenesis and different types of transposon mutagenesis.

UNIT II       

          Transposition – definition  – process of transposition

Transposable genetic element  - Insertion sequences - Simple transposition modules.

Composite transposons  - IS modules.

UNIT III

           Replicative mechanism of transposon

 Transposition by both replicative and non-replicative mechanisms.

 Transposase and resolvase.

UNIT IV

             Detailed description of

Ψ     Tn 10,         Tn 5

Ψ     Tn 9      Tn 3

UNIT V

            Applications of transposons in:

Ψ     Agriculture,

Ψ     Medical, 

Ψ     Industrial research.

 

PAPER 6: GENE EXPRESSION

UNIT I

Ψ     Gene expression in eukaryotes  

Ψ     Enhancers activate gene expression over long distances

Ψ     Genes can be transcribed in cell free extract

Ψ     Transcription factor are purified and cloned

UNIT II

Ψ     Cloning in Strepomyces species

Ψ     Cloning in Saccharomyces cerevisiae

Ψ     Development of vectors 

Ψ     Choice of vector for cloning

Ψ     Expression of cloned genes in yeast.

UNIT III

Ψ    Eukaryotic vectors - SV40 and its properties –

Ψ    Experiments with late region replacement

Ψ    Construction of improved SV40 vector

Ψ    Lambda phage vector and its properties

UNIT IV

Ψ    Specialized cloning vectors for cDNA

Ψ    Phages

Ψ    Cosmids      &        Phasmids

Ψ    Synthesis of specific RNA in in-vitro

UNIT V

Ψ     Selection of vectors for copy number

Ψ     Plasmid - Conjugative- Non Conjugative- Low copy number - High copy number -Degradative plasmid –Ti plasmids. Cloning Promoters, and Terminators.

 

PAPER 7: Plant & ANIMAL biotechnology

UNIT I

1.Plant cloning vectors

Ψ     Agrobacterium tumefaciens

Ψ     Ti plasmids of Agrobacterium

Ψ     Derivatives of Ti plasmids

Ψ     Viruses as vectors for whole plants

Ψ     T – DNA modified to act as a gene vector

Ψ     Transgene expression in plant tissues

Ψ     Plant vaccines

UNIT II

1.  Animal cell cloning vectors

Ψ     Baculoviruses

Ψ     Bovine papilloma virus

Ψ     Adenovirus

UNIT III

1.  Mammalian expression vectors

Ψ     Retroviral vectors

2.  Gene expression analysis:

Ψ     SDS - PAGE

Ψ     Western Blotting

Ψ     Southern Blotting

Ψ     Northern Blotting

UNIT IV

1.  DNA sequencing:

Ψ     Maxam Gilbert method of sequencing

Ψ     Sanger’s dideoxy nucleotide method

2.  RFLP

Ψ     Principle of RFLP

Ψ     Application of RFLP

Ψ     Bacterial restriction endo-nucleases digestion analysis

UNIT V

1.  DNA fingerprinting

Ψ     Steps involved in DNA fingerprinting

Ψ     Application

2.  PCR

Ψ     Principle

Ψ     Standard PCR

Ψ     Single sided PCR

Ψ     RT –PCR

Ψ     Random PCR

Ψ     Inverse PCR.

 Practical-1  

  1. Familiarizing with the lab and equipments
  2. Prepare individual reagents, collect materials, prepare solutions, and pour plates.
  3. Measurements, math and micropipettor practice.
  4. Bacterial culture techniques Streak for colony isolation.  Set up liquid Overnight culture.
  5. Growth curve study
  6. Count colonies
  7. Genomic DNA extractions.  Bacterial DNA prep
  8. Genomic DNA extractions. Fungal DNA prep.
  9. Genomic DNA extractions. Algal DNA prep
  10. Plasmid DNA extraction
  11. Agarose gel electrophoresis of genomic DNA
  12. Agarose gel electrophoresis of plasmid DNA  
  13. Cloning of DNA into expression vectors
  14. Single cell electrophoresis to assess DNA damage  
  15. Transformation of microbes  
  16. Preparation of competent cells 
  17. Induced mutagenesis and isolation of antibiotic resistant and auxotrophic strains  
  18. Single colony isolation and checking for markers 

Practical-2  

  1. Restriction digestion of Bacterial genomic DNA and analysis on Agarose gel       electrophoresis
  2. Restriction digestion of Fungal / algal genomic DNA and analysis on Agarose gel electrophoresis
  3. Restriction digestion of plasmid DNA and analysis on polyacrylamide gel electrophoresis
  4. Restriction fragment length polymorphism (RFLP)
  5. Purify DNA fragments for cloning
  6. Gene amplification (PCR)
  7. Experiments with gene fusion
  8. Analyze DNA ligations by gel electrophoresis
  9. Transform competent bacteria (chemical poration / electro poration)
  10. Conjugation studies
  11. Coliphage isolation
  12. Transduction trials in E. coli
  13. Production of proteins and analysis by SDS-PAGE.
  14. Western blot / Immunoblot
  15. Southern hybridization
  16. Transposon mutagenesis of chromosomal DNA
  17. Site directed mutagenesis
  18. Use of non-radioactive probes

      

Reference Books:  

1.   Molecular cloning – J.Sambrook, E.F.Fritsch and T.Maniatis.
 2.   Gene cloning – T.A. Brown
3.   Principles of gene manipulation – Old & Primrose  
4.   Manipulation and expression of r DNA –  Domnique Robertson, Scottshore,        David M. Miller
5.   Vector – A survey of molecular cloning vector and their uses- Raymond Rodriguez and David
      T. Denhart  
6.   From genes to clones – Introduction to gene technology – Errst – L. Winnacker 7.   Gene expression
      technology: Methods in enzymology (Vol.185) – David V. Goeddel 
 8.   Methods in Gene Biotechnology – William Wu, Michael J.  Welsh, Peter B. Kufrmar, Helen H. Zhang
 9.   Maximizing gene expression – William Rezmikff, , Lamy   Gold
10. Molecular genetics of Bacteria – J.W. Dale 
11. Principles of genetics – Gardner, Simmons and Snustad  
12. Gene VII – Benjamin Leuvin  
13. Basic genetics – D.L.H. Hartl  

  • THE COURSE IS OF 2 SEMESTERS - EACH 6 (SIX) MONTHS DURATION. 

  • FEE : Rs. 50,000/- (FIFTY THOUSAND) PER SEMESTER Excluding Hostel fee.

  • Placement assistance

  • educational loan ASSISTANCE

  • abroad placement ASSISTANCE

  • GRE/TOEFL EXAMS ASSISTANCE

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2.  DIPLOMA IN Immunotechnology- 2009-10

       Introduction: - Immunotechnology has grown to encompass many fields such as allergy, clinical  immunology,  immunochemistry,  immunopathology,  immuno pharmacology, tumor immunology and transplantation immunology.  Immunology has always depended upon and  stimulated  the application of technology such as the  use  of  microscopy,  electrophoresis,  radiolabelling,   Immunofluorescence, recombinant  DNA,  transgenic animals, monoclonal antibodies, animal cell/tissue culture, vaccine development and plantibodies production.  The modern medicine is  the  translation  of  advances  in  immunochemistry  and  immunobiology   into diagnostic and therapeutic procedures, that will be useful and very effective in the generation of many skilled job opportunities in all the above fields mentioned.

Paper I – General Paper

Unit I  : Life Skills & Communications:

01)        a. External and internal influences in one’s life.

02)        a . Meaning and process of coping

             b. Mechanism of coping

             c. Coping with physical change and sexuality

03)        a. coping with shyness, fear

             b.Coping with anger, criticism , failure

04)         a. Self-Acceptance, Self-esteem

             b.Self-actualization

05)        a. Positive thinking- motivation

             b. Goal setting

06)        a. Problem solving

             b. Decision making

07)        a. Time Management

             b. Stress Management

08)        a. Leadership

             b. Team building

09)        a. Inter personal elements of communication

             b. Expression. Listening and responding

10)        a. Group Communication and Group discussion

 

Books for Reference

  0  1)      “ Organisational Behaviour” by Jit S.Chandan

      2)      “Organisation Behaviours” by M.M.Varma and Adarval

      3)      “ How to be your own best friend” by Paul Mauck

 

  

Unit II  : Spoken English:  

S

Situational Skills:

Greeting – Introducing oneself –Inviting, someone – Making requests Offering help –   Seeking permission –    Asking for advice – Expressing gratitude –Asking about remembering-   – persuading  - complimenting / Congratulating – Expressing sympathy – complaining – Apologizing – Making suggestions – Warning someone – Starting conversation with a stranger – Leaving someone for a short time – Ending a conversation – Asking for information – Asking for someone’s Opinion – Asking if someone’s sure –Asking someone to say something again checking that you have understood – Asking whether someone knows – Asking about  possibility –   Asking about preference – Asking if someone     is able to do something – Describing something – Some   useful expressions.   Building Vocabulary: One word substitution  - Synonyms - Homophones – Odd man out. Language Use: Identification of tenses and agreement of verbs. Statements, Negatives,  Interrogatives and Question tags. Rectification of errors in articles. Propositions and adverbs  – Comprehension – telegraphic Message. 

 

Books for Reference:

 

 01) Spoken English for  you – Emerald Publishers – Authors:

           G.Radhakrishna Pillai ,K.Rajeevan.

 02)  Spoken English  - A Self Learing  Guide to Conversation Practice      

      Authors: V.Sasikumar, P.V.Dhamija

 

Unit III : Computer Usage and Applications:

01)    Introduction to Computer: Generation and History of Computers -Characteristics of Computer – Digital Computer - Analog -          Computer –     Hybrid Computer. Classification of Digital Computer: Micro Computer –Mini Computer – Mainframe computer

02)    Input Devices: Keyboard, Mouse, Light pen –Output Devices: Monitor, Classification of Monitors, Printers, Classification of printers –Study of    Secondary storage devices.

 03)   Hardware, Software, farmware, parts of CPU: ALU – Control Unit –  Main   memory – RAM, ROM.

 04)   System Software: Interpreter, Assembler, Compilers – Operating Systems: DOS – Booting System – Internal and   External Commands – Introduction to High-level languages.  

 05)   Origin of Internet – Appanet – MODEM – ISP (VSNL) –  Upload, Download. E-mail: Origin of Worldwide web (WWW) –   Browsers – HTML     Basics.  

      Books for reference:

01)   Introduction to Computers by Radhakrishnan.  

02)  Fundamentals of Computers by V. Raja Raman.

Paper 2: Immune system

Unit I: Types of Immunity

Antigen:  Essential feature of antigens - Factors that influence

      Antigenicity – Epitopes - Haptens          

 a)     Innate immunity   

1.     Anatomic barriers

2.     Physiologic barriers (Lysozyme, Interferons)

3.     Endocytic and Phagocytic barriers (PMN & Macrophage)

4.     Barriers created by Inflammatory response.

       b)    Acquired Immunity –

1.     Cells of immune system (B & T cells)

2.     APC (Antigen presenting cells)   

3.     Functions of Humoral and Cell mediated Immune response

             Unit II: Anatomy of Lymphoid Organs.

a) Source of lymphoid cells –Bone marrow

            b) Primary lymphoid Organs

              1.Thymus – Thymocytes - Skin - Bursa of Fabricius - Payer’s patches

c). Secondary lymphoid organs

              Lymph nodes – Structures – Spleen - Bone marrow       

 u     Unit III: Immunoglobulin Structure and Function

   Basic structure and functions of Immunoglobulin

  IgG  -  IgA  -  IgM  -  IgD -   IgE

     Unit IV: Memory cells

1.  Generation of Memory cells and Plasma cells

2.  Memory T & B cells

3.  Idiotypes

 

Unit V: Lymphocyte Differentiation

 T lymphocytes – B lymphocytes

Paper 3: Positive and Negative Antigen and Antibody interaction

 Unit I: Biology of complement system

             Definition of complement - Components of complement - Types of Complement Pathways – Classical – Alternate - Regulation of complement system - Biological constituents of complement activation  - cell lysis - Inflammatory response – Opsonisation- Viral neutralization  - Solubilisation of Immune complexes - Complement deficiencies - Structure and Function of MHC –I & II. – Structure - Gene arrangement – Polymorphism - Function

Unit II : a). Antigen recognition and presentation

 Endogenous – Cytosolic pathway - Exogenous – Endocytic pathway - Self MHC restriction of T-cells - Cells that function in Antigen presentation – TAPS Transporter Associated with Antigen Processing

 b). Cellular Mediated Immunity

Helper T cells - Effector T-cell function - Complement Mediated cell lysis

 Unit III: Hypersensitivity reactions

1. Type I Hypersensitivity or Anaphylactic reaction Phenomenon of Anaphylaxis - Atopic allergy - Food allergy

2. Type II Hypersensitivity or ADCH  Transfusion reaction - Rh factor - Organ transplants –

3. Type III Hypersensitivity or Immune complexes Arthus reaction - Reaction to internal and inhaled antigen - Serum sickness -

4. Type IV Hypersensitivity or Cell mediated immunity.

Unit IV: Autoimmune Disorders

1.Scope of Autoimmune diseases    2.Physiology of autoimmunity

3.Types of Autoimmunity

a. Organ specific autoimmune disease (Myasthenia gravis)

b. Systemic autoimmune disease (Systemic Lupus Erythematous)

4.Mechanism for induction of autoimmunity  

Unit V  a). Tissue Grafting

Genetic control of transplantation - Mechanism of graft rejection  - Prevention of graft rejection

i. Tissue typing ii. General immunosuppressive therapy iii. Specific immunosuppressive therapy

b). Transplantation strategies

 1.Bone marrow              2.Organ transplantation

c). Graft Vs Host Disease

1.Activation and Proliferation of cells  

Paper 4:           Analytical methods of immune system

Unit I
          Antigen Isolation  - Sonication – Lyophilization – Thermal extraction methods – TCA method – Ethanol extraction – Triton x 100 extractions – isolation of membrane bound proteins – cytosolic proteins – flagellar proteins – enzymes.
Unit II

       Purification and characterization of various antigens - Ammonium sulphate liquefaction – salting in – salting out – Chromatographic techniques – gel exclusion – ion exchange – NMR - Haptens from pathogens and other biological molecules.

Unit III     

     Biophysical and chemical and affinity separation methods – Paper chromatography – Thin layer chromatography - HPLC – GLC – Electrophoretic techniques – SDS- PAGE – cellulose strip electrophoresis - Centrifugation – density gradient centrifugation – Isopycnic – Ultracentrifugation.

Unit IV

        Production of antibodies – adjuvants – Freunds incomplete & complete adjuvants – Selection of experimental animals and their uses – Restrain and handling of experimental animals - vaccination schedule.

Unit V

          Purification of antibodies - Quantification for Immunoglobulin by RID, EID and Nephlometry – Specificity of antibodies – CIE and WB methods.

Paper 5: Monoclonal antibody production
Unit I

          Hybridoma and monoclonal antibody production : Animal Cell cultures – cell line maintenance – cell fusion – myeloma cells – technique of monoclonal antibody production - genetically engineered antibodies – fused hybridomas.

Unit II

          Immuno-diagnosis:  precipitation assays – Mancini’s single dimension assay – ODD – RID – Direct agglutination – Indirect (passive) agglutination – hemagluttination – Coombs test – Bentonite Flocculation test – Rose-Waaler test - Macroscopic slide agglutination – microscopic agglutination test (MAT), latex agglutination assays – RIA - ELISA – Biotin-avidin enhanced immunoassays - Complement fixation test; assessment of immune complexes in tissues.

Unit III

        Application of monoclonal antibodies in biomedical research – Enumeration of human lymphocyte sub population – Cell depletion – Cell isolation – Probing function of cell surface molecules – diagnosis in cancer – antitumor therapy by drug delivery system - imaging – nephlometric assays  - analysis of complex antigen mixture – purification of antigens – analysis of embryological relationship – monoclonal mutants.

Unit IV

          Human monoclonal antibodies – Human antimouse antibodies – MHC linked target of cytotoxic T cells -  Catalytic antibodies – HLA antigen detection – viral detection and sub typing – parasitic identification – Immuno suppression - fertility control.

 

           Paper 6: Cell mediated immunity assay and application

 Unit I

Purification of mononuclear cells from peripheral blood - Isolation and characterization of T-cell subsets Helper – suppressor cell ratio – E Rosette-forming cells– human T cell specific markers – T cell subsets – Production of T cell antibodies – detection of T cell antigens with specific antisera -  Lymphocyte assays - B-cells and macrophages – Human B cell specific markers – Surface immunoglobulins – Cytoplasmic immunoglobulins.

 Unit II

Fluorescent activated cell sorter (FACS) – cell selection by FACS – Immunofluorescence techniques – Direct test with labeled antibodies – Indirect test for antibodies – High resolution with the confocal microscope – Flow cytometry – Flow cytofluorimetry - Mitogen and antigen induced lympho proliferation assay.

 Unit III

Mixed lymphocyte reaction – mixed lymphocyte culture  - Cell mediated lympholysis – Clinical application of T & B cell assays -           Assessment of delayed hypersensitivity reaction - Macrophage culture -           Assay for macrophage activation -            enumeration of macrophages – macrophage role in host defense, human immunodeficiency virus, interferon gamma, interleukin – 2, multiple sclerosis, phagocytic responses, secretory activity - dendritic cells - Isolation and role in human.

 Unit IV

In-situ & In-vivo characterization of cells from tissues : Nucleic acid probes – hybridization assays – Southern blot – Generation of T-cell clones - Gene Rearrangement assay for lymphocyte clonality – In-situ hybridization – target amplification technique (PCR) – methods of analyzing RNA - HLA typing

            Paper 7 :         Vaccine development technology

 Unit I

Biology & Assay of cytokines : Interleukin –1 and tumor necrosis factor – properties of human interleukins and other immunoregulatory cytokines – target cells IL-1 and TNF - actions of IL-1 and TNF on hematopoietic and lymphoid and non-lymphoid cells and tissues – IL-1,TNF & their inhibitors as therapeutic agents – IL-2 – receptor and signal Transduction – role of IL-2in human immune system – IL-6 and related cytokines – antiviral interferons – immune interferons –Transforming growth factor β.

Unit II

Vaccine technology including DNA vaccines: Acquired memory –experiment vaccines in development – malaria – cholera – TB – Schistosomiasis – Adjuvant – Killed organisms as vaccine – Live attenuated organisms as vaccines – Subunit vaccines – protein vaccines DNA vaccines - naked gene acting as a vaccine – rDNA Vaccines.

Unit III

Identification of T & B epitopes for vaccine development – autoimmunity – T – down regulation – Original antigenic sin - current vaccines and their development.

Unit IV
Immunotechnology and infectious diseases: Diagnosis of infectious hepatitis – infectious mononucleosis – immunoblastic reaction – infectious rhinitis – infectious syndrome – infective endocarditis – immune therapies.

Unit V
Immunity and genetic engineering in transgenic animals – immunoassays for bacteriophage λ library screening – other gene libraries screening – screening for monoclonal antibody production - Immunoscreening of recombinant library.

 
Practical I

Immunity to pathogens and raising Polyclonal antibodies 
Vaccine development 
Mancini’s single dimension diffusion test 
ODD
Rocket electrophoresis 
CFT 
 HA 
MAT 
Serum electrophoresis 
Immune electrophoresis 
CIE 
ELISA 
Western/Immunoblotting assays 
Immunofluorescence assays.

Practical II

Technology of hybridoma and monoclonal antibodies   
Culture and maintenance of myeloma cell lines   
Cell fusion   
B cell and T cell isolation   
Monoclonal antibody in biomedical research practice   
Application of human monoclonal antibodies   
T-cells biotechnology  
T cell cloning   
Lymphokine production  
Role in immunological reactions and applications in vaccine development.

 

Reference Books

 

1.   Immunology – I.Roitt, J.Brestoff and D.K.Male

2.   Medical Immunology – Gabriel Virella

3.   Basic & Clinical Immunology - Daniel P.Stites., Abba I.Terr & Tristram G. Parslow

4.   Diagnostic Immunology – Keren & Warren

5.   Principles of Immunology – I.Roitt

6.   Immunological Techniques – D.M.Weir

7.   Current protocols in immunology – 3 Volumes – Wiley Pub. 1994.

8.   Immunology – Ian R. Tizard

9.   Immunology – Janis Kuby  

        FEE : Rs. 50,000/- (FIFTY THOUSAND) PER SEMESTER

                   Excluding hostel fee.

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DETAILS OF Short TERM Courses

 1.  Cloning and Expression -  2009-10

This course assumes participants have a basic understanding of recombinant DNA techniques and have some basic lab skills such as pipetting and reasonable lab math skills. Lecture and discussion will cover cloning strategies and selection of appropriate hosts, vectors and enzymatic tools, with an emphasis on how cloning and expression techniques would be applied in typical industry settings. In conjunction with the lecture, students will undertake a cloning/expression project. This project will give students many basic skills commonly used in the molecular biology lab.

Course Format:

Eight classes/labs that last for 4 hours each. Each class/lab will begin with a lecture and outline goals of each lab. In some instances the lab will predominate class time and vice versa. In many instances we will have prolonged breaks due to lab design (incubations), which will be filled in by demonstrations and lectures. Students will work in pairs within the lab. Students must supply own lab coat and safety glasses.

Week 1

Class 1

Lecture: Introduction

· Outline goals of this course both lecture series and the lab component.

· Define cloning and expression

· Overview tools of cloning

o Enzymes

o Vectors

o Hosts

Lab: Introduction

· Familiarize ourselves with the lab and equipment.

· Overview of basic molecular biology techniques.

· Prepare DNA samples for cloning experiment

Class 2

Lecture: Where do we begin the cloning process?

· Types of libraries- cDNA, cDNA expression, phage display, two hybrid.

· Screening libraries by:

o DNA hybridization

o PCR

o Expression

Lab: Clone LiP  into pBR322

· Analyze DNA digests from last lab

· Purify DNA fragments for cloning

· Ligate LiP DNA to expression vector.

Week 2

Class 3

Lecture: Libraries and Screening continued:

· Libraries in Yeast

o functional screens in yeast-complementation

o Two hybrid screen/genomic two hybrid screen

Lab: Monitor ligations

· Analyze DNA ligations by gel electrophoresis

· Transform competent bacteria.

Class 4

Lecture: Clone characterization

· sub cloning

· sequencing

· Data base analysis

Protein expression

· Applications of recombinant protein

· Protein expression in vitro

· Protein expression in bacteria

o Inducible systems to minimize recombinant protein toxicity

o Soluble/insoluble proteins -inclusion bodies

Lab: Assess transformation

Set up overnight cultures of potential clones

Week 3

Class 5

Lecture: Tagging systems for detection and purification

· Native versus tagged proteins.

· Why tag proteins?

· Tagging systems.

· How tags work in protein purification

Lab: Determine whether your cloning was a success

· Perform minipreps on potential clones

· Analyze clones by restriction digest and gel electrophoresis

Week 4

Class 6

Lecture: Protein expression in eukaryotes:

Vectors: plasmid or virus based.

Purpose:

1) Identify regulatory sequences in mammalian genes.

2) Protein localization

3) Over expression for biochemical and phenotypic studies

4) Over expression for purification

Systems:

1) Yeast

2) Insect cells

3) Mammalian cells

Lab: Mini-inductions

· Induce expression of your new recombinant protein and monitor expression by

microscopy and SDS-PAGE

Class 7

Lecture: Protein expression in eukaryotes: continued

New technology in vector construction:

· One step cloning systems:

o Stratagene: Exchanger system

o Invitrogen: Gateway technology, Echo technology

o Clontech: Creator system

Application of recombinant proteins

· Immunization

· Protein replacement therapy

· Protein structure

o drug design

o protein function

· Limitations of recombinant proteins

Lab: Large scale induction to prepare and purify LiP fusion protein.

Class 8

Lecture: RNAi, cloning a mouse

· Assess the function of your newly cloned gene product– the next step in

understanding the function of your gene in development and disease.

· Cloning other mammals- pros and cons

Lab: Purify LiP protein and analyze by SDS-PAGE.

TRAINING FEE : 200 US $ / RUPEES 10,000/- (TEN THOUSAND ONLY) Excluding Hostel fee.

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2.  PLANT AND ANIMAL TISSUE CULTURE 2009-10

SYLLABUS AND SCHEDULE

Class is typically: 7 Sessions of 2.5 Hours Each

COURSE DESCRIPTION: 

       This course will investigate the fundamentals of cell and tissue culture in animal and plant systems. Participants will become familiar with sterile technique and media preparation. Covered topics include transformation and regeneration of plants from culture. In animal cell culture, topics include subculture and viability staining.

COURSE FORMAT: This course will include an introduction to theoretical aspects of plant and animal culture with emphasis on practical application. Laboratory exercises will be the primary focus of the course. Students will be responsible for reading and discussing current literature in order to become familiar with the utility of these techniques in biotechnology.

SCHEDULE:

Preparation:       Regeneration of shoots and roots from callus in culture takes approximately ten weeks, so for the students to be able to observe plant regeneration it is necessary to have callus tissue growing for no less than one month, and ideally six weeks prior to the first day of the course.

Session 1:

Lecture: Plant Tissue Culture and Regeneration Overview

Lab exercises:

1. Transfer callus to fresh media

2. Media preparation for Session 2.

Session 2:

Lecture: Agrobacterium tumefaciens mediated transformation

Lab exercises:

1. "Floral dip" method of Arabidopsis transformation

2. Growing callus derived from African violet petiole explants in culture Preparation and plating of explants

Session 3:

Lecture: Animal tissue culture fundamentals: growth conditions, biological safety cabinets, and media, adherent vs. suspension cultures, tissue culture formats.

Lab Exercise: Set up CEF cultures for continuous growth. Set up Zeocin antibiotic kill curves. (We will maintain these cultures throughout the month)

Session 4:

Lecture: Tissue culture contaminants focusing on mycoplasma.

Lab Exercise: Harvest K562 and SKBR3 from session 3 to look at effects of antibiotics. (More practice with subculture and viability staining.) Test cells and cell supernatants for mycoplasma infection.

Session 5:

Discussion: Agrobacterium-mediated transformation

Lab exercise: Screening for transformants (the GUS reporter gene)

Transfer shoots to rooting medium (high auxin)

Session 6:

Lecture: Primary tissue cell culture; cloning schemes and options for picking clones

Lab Exercise: Analyze mycoplasma results. Set up cloning schemes and plates using EL-4/GFP or K562/luc.

Session 7:

Lecture: Transfection; hybridomas; cryopreservation

Lab Exercise: Pick clones. Analyze data showing alternative methods from visual clone picking.

HANDOUTS

Clough, Steven J and Andrew F. Bent (1998) Floral dip: a simplified method for

Agrobacterium-mediated transformation of Arabidopsis thaliana. The Plant Journal 16, 735-743.

Peter R. Day (1996) Genetic modification of plants: significant issues and hurdles to success. American Journal of Clinical Nutrition 63, 651S-656S.

Gottschalk, S., Edwards, O., Uluhan, S., Huls, M., Goltsova, T., Davis, A., Heslop, H., and C. Rooney (2003) Generating CTLs against the subdominant Epstein-Barr virus LMP1 antigen for the adoptive immunotherapy of EBV-associated malignancies. Blood 101, 1905-1912.

Notice to Students: This information is for guidance only. The actual content of your class may differ as the instructor chooses.

TRAINING FEE : 200 US $ / RUPEES 10,000/- (TEN THOUSAND ONLY) Excluding Hostel fee.

 3.  Immunotechnology Lab  Course -  2009-10
    SYLLABUS AND SCHEDULE

The goal of this laboratory is to provide an experience working with Immunochemical techniques. The laboratory will have several sections that will be outlined below. In addition to the new techniques you will be exposed to this quarter, a major component of the lab will be to allow you to take part in the design of experiments. There will be no assigned laboratory manual. We will use reference books in the lab as well as web resources to find protocols to achieve to goals of the lab. 

Week Topic Techniques

1 Introduction

Experimental planning

2-4 Antibody purification and labeling

- Ammonium Sulfate precipitation and dialysis

- Quantification of protein concentration

- DEAE fractionation of IgG and IgM

- PAGE analysis of fractions (reducing and non-reducing)

- Affinity purification of antigen specific antibodies

- Biotin Labeling of Ig

5-7 Use of Labeled antibodies

- Dot Blots

- Western Blots

- ELISA

8-9 Use of antibodies to isolate proteins and protein complexes

- Immunoprecipitation and co-IPs

 TRAINING FEE : 200 US $ / RUPEES 10,000/- (TEN THOUSAND ONLY)  
 Excluding Hostel fee.

 

. To know about the Training Methodologies                                          

               Recently we have started in-plant training programs for B.E / B.Tech. students in the fields such as Biotechnology, Molecular Biology, Genetic Engineering and Bioinformatics.  Summer training programs are also organized for not only engineering graduates but also for graduates and postgraduates of Science faculties.

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