BSc Bio -Tech THRID YEAR-

Molecular biology and r-DNA technology

UNIT – I Gene expression
1.1 . Transcription in prokaryotes: Enzymatic Synthesis of RNA, Basic features of RNA synthesis, E.coli RNA polymerase, Classes of RNA molecules, Transcription mechanism in prokaryotes- Promoter, initiation, elongation, proof reading and Rho dependent and Rho independent termination.
1.2 . Transcription in Eukaryotes: Polymerases of eukaryotes, Promoters of eukaryotes,
1.3 . Synthesis of hn RNA, Splicing mechanisms-Self splicing, protein mediated splicing, alternative splicing, Capping and polyadenylation.
1.4 . The Genetic Code, properties of genetic code, Wobble hypothesis.
1.5 . Translation mechanism in prokaryotes and eukaryotes

UNIT – II Regulation of Gene expression
2.1. Regulation in Prokaryotes: General aspects of Regulation
2.2. Transcription level regulation-positive, negative, auto and coordinated regulation
2.3. Operon concept – lac, trp, operons.
2.4. Transcriptional Control through Transcription factors.
2.5 .Translation regulation in Eukaryotic and prokaryotic organism

UNIT – III r-DNA technology
3.1. Enzymes used in gene cloning: restriction endonucleases, ligases, phosphatases, methylases, kinases.
3.2. Cloning vehicles, plasmids, cosmids, phage vectors, Shuttle vectors Baculovirus vector system, expression vectors, expression cassettes.
3.3. Construction of genomic and cDNA libraries. Identification of cloned genes
3.4. Principles involved in blotting techniques- southern, northern and western.
3.5. Principles and application of PCR Technology and DNA fingerprinting technique and its application.

REFERENCE BOOKS:

  1. Molecular Biology of the Gene – By Watson, Hopkins, Goberts, Steitz and Weiner (Pearson Education)
  2. Cell and Molecular Biology – By Robertis & Robertis, Publ: Waverly
  3. Text Book of Biotechnology – By H.K. Das (Wiley Publications)
  4. Gene Structure & Expression – By J.D. Howkins, Publ: Cambridge
  5. Genetic Engineering – By R. Williamson, Publ: Academic Press
  6. Principles of Gene Manipulation – By R.W. Old & S.B. Primrose, Publ: Blackwell
  7. Genes – By B. Lewin – Oxford Univ. Press
  8. Molecular Biology & Biotechnol. – By H.D. Kumar, Publ: Vikas
  9. Methods for General & Molecular Bacteriology – By P. Gerhardf et al., Publ: ASM
  10. Molecular Biotechnology – By G.R. Click and J.J. Pasternak, Publ: Panima
  11. Genes and Genomes – By Maxine Singer and Paul Berg
  12. Molecular Biology – By D. Freifelder, Publ: Narosa

ELECTIVE PAPER-(A) PLANT BIOTECHNOLOGY

UNIT – I: Introduction
1.1. Historical perspectives of plant tissue culture, and Basic requirement for tissue culture laboratory
1.2 . Culture mediums for plant tissue culture- MS medium and B5 Medium. Sterilization of media-steam, dry and filter sterilization- Explants sterilization
1.3. Plant growth regulators and differentiation.
1.4.Method of tissue culture-formulation of medium explants collection, surface sterilization, inoculation, Callus induction, subculture and regeneration of plants
1,5. Suspension cultures- growth and subculture, types and synchronization of suspension cultures.

UNIT – II: Applications of plant tissue culture
2.1. Meristem culture and its uses in production of virus free plants
2.2. Clonal propagation, Micro propagation of plants – medicinal plants and endangered plants –method and advantages
2.3. Somatic embryogenesis- Principle, protocol and importance. Artificial seeds – production, applications and limitations.
2.4. Anther culture and production of androgenic haploids.
2.5. Somaclonal variations; – sources of somaclonal variatins, selection of soma clones, progeny testing of soma clones, applications of somaclonal variations to crop improvement, Embryo rescue

UNIT – III: Applications of plant tissue culture
3.1. Protoplast – properties of protoplast ,Protoplast – Isolation (mechanical and enzymatic methods ), Culturing and regeneration of protoplasts , Different methods of protoplast fusion (mechanical fusion, chemo fusion, electro fusion) and Selection of somatic hybrids and cybrids.
3.2. Cryopreservation of plant cultures and application of plant tissue culture .
3.3. Immobilization of cells and the effect of elicitors on the production of secondary metabolites of commercial value
3.4. Introduction to Agrobacterium tumifaciens, Features of Ti Plasmid, molecular mechanism of T-DNA transfer.
3.5. Physical gene transfer methods – Particle Bombardment, Electrophoration and Microinjection.

REFERENCE BOOKS:

  1. Plant Tissue Culture and its Biotechnological Applications By W. Barz, E. Reinhard, M.H. Zenk
  2. Plant Tissue Culture By Akio Fujiwara
  3. Frontiers of Plant Tissue Culture By Trevor A. Thorpe
  4. In vitro Haploid Production in Higher Plants by S. Mohan Jain, S.K. Sopory, R.E. Veilleux
  5. Plant Tissue Culture: Theory and Practice By S.S. Bhojwani and A. Razdan
  6. Plant Cell, Tissue and Organ Culture, Applied and Fundamental Aspects By Y.P.S. Bajaj and A. Reinhard

ELECTIVE THEORY (B) MEDICAL BIOTECHNOLOGY

Unit 1: Methods for diagnosis of human diseases
1.1 Karyotyping of human chromosomes
1.2 Chromosome banding– G banding and R-banding technique
1.3 Inheritance patterns in Man– Pedigree analysis
1.4 Diagnosis using monoclonal antibodies- ELISA
1.5 DNA/RNA based diagnosis– HBV, HIV

Unit 2: Inherited disorders
2.1 Chromosomal disorders caused due to structural chromosomal abnormalities (Deletions, duplications, Translocations)
2.2 Chromosomal disorders caused due to numerical chromosomal abnormalities (autosomal and allosomal)
2.3 Monogenic disorders (autosomal and X-linked diseases)
2.4 Mitochondrial diseases – LHON, MERRF
2.5 Cancer – types, molecular basis of colon cancer and breast cancer

Unit 3: Therapeutic approaches for human diseases
3.1 Gene therapy – exvivo and invivo gene therapy; somatic and germline gene therapy;
3.2 Strategies of gene therapy: gene augmentation – ADA deficiency; Prodrug therapy / suicide gene – glioma
3.3 Stem cells – potency definitions; embryonic and adult stem cells; applications of stem cells – cell based therapies and regenerative medicine
3.4 Encapsulation technology and therapeutics-Diabetes
3.5 DNA based vaccines, subunit vaccines – Herpes Simplex Virus, Recombinant attenuated vaccines– Cholera

REFERENCE BOOKS:

  1. Medical Biotechnology-Pratibha Nallari, V.Venugopal Rao-Oxford Press
  2. Introduction to Human Molecular Genetics – J.J Pasternak, John Wiley Publishers
  3. Human Molecular Genetics –Tom Strachen and A P Read, Bios Scxientific Publishers
  4. Human Genetics Molecular Evolution, Mc Conkey
  5. Recombinant DNA Technology, AEH Emery
  6. Principles and Practice of Medical Genetics, I, II, III Volumes by AEH Edts. Emery
  7. Molecular Biotechnology, Glick and Pasternak

GENERIC ELECTIVE AGRITECHNOLOGY

Unit I GM Crops:

  1. Crop evolution and human civilization; Introduction to GM Crops
  2. Applications of GM Crops, : (Bt- Cotton; and golden rice), Global area of biotech crops
  3. Implications of GM-Crops in Agriculture, Public perception on GM crops and scientific solutions.
  4. Bioethical issue on GM crops
  5. Crop seasons and crop rotation, Sustainable agriculture; pheromones, traps etc

Unit II Organic farming and Conservation

  1. Introduction to organic farming, green manure production, Soil fertility and management
  2. Role of earthworms in soil structure, and productivity, Cost-benefit analysis of vermicomposting
  3. Invitro propagation and Conservation of forest and medicinal plants
  4. Conservation of germ plasm, Indigenous cultivars & wild plants
  5. Commercial cultivation of medicinal and aromatic plants

REFERENCE BOOKS:

  1. Plant Tissue Culture By Akio Fujiwara
  2. Frontiers of Plant Tissue Culture By Trevor A. Thorpe
  3. Plant Cell, Tissue and Organ Culture, Applied and Fundamental Aspects By Y.P.S. Bajaj and A. Reinhard
  4. Text Book of Biotechnology – By H.K. Das (Wiley Publications)
  5. Biotechnology – By H.J. Rehm and G. Reed. VIH Publications, Germany
  6. Biogas Technology – By B.T. Nijaguna
  7. Biotechnology – By K. Trehan
  8. Food Microbiology – By M.R. Adams and M.O. Moss
  9. Introduction to Biotechnology – By P.K. Gupta
  10. Essentials of Biotechnology for Students – By Satya N. Das

MICROBIAL TECHNOLOGY

Unit 1: Introduction to Microbial technology
1.1. Introduction to industrial biotechnology, scope and applications
1.2. Principles and exploitation of microorganisms and their products
1.3. Isolation and screening of microorganisms for industrial products
1.4. Strategies for Strain improvement (mutation, selection, recombination)
1.5. Preservation of industrial microorganisms

Unit 2: Microbial fermentation
2.1 Principles of Fermentation technology
2.2 Fermentation concept and Design
2.3 Types of Fermentations, Formulation and Design of fermentation Media
2.4 Substrates used as Carbon and Nitrogen Inoculum development.
2.5 Factors affecting fermentation process

Unit 3: Microbial technology products and applications
3.1 Microbial production of Organic acids (Lactic acid and citric acid)
3.2 Microbial production of Amino acids (Glutamic acid and Aspartic acid)
3.3 Fermentation by microbes for food additives: dairy products (Bread and SCP), beverages (Beer and Wine) and antibiotics (Penicillin and Streptomycin)
3.4 Therapeutic drugs: Monoclonal antibodies and insulin,
3.5 Biofuel: Alcohol and Methane

REFERENCE BOOKS:

  1. Text Book of Biotechnology – By H.K. Das (Wiley Publications)
  2. Biotechnology -By H.J. Rehm and G. Reed. VIH Publications, Germany
  3. Biogas Technology – By b.T. Nijaguna
  4. Biotechnology – By K. Trehan
  5. Industrial Microbiology – By L.E. Casida
  6. Food Microbiology – By M.R. Adams and M.O. Moss
  7. Introduction to Biotechnology – By P.K. Gupta
  8. Essentials of Biotechnology for Students – By Satya N. Das
  9. Bioprocess Engineering – By Shuler (Pearson Education)
  10. Essentials of Biotechnology – By Irfan Ali Khan and AtiyaKhanum (Ukaaz Publications)

ELECTIVE PAPER- (A) ANIMAL BIOTECHNOLOGY

UNIT-I Introduction to Animal Biotechnology
1.1. Animal tissue culture, history, requirements for animal cell culture
1.2. Substrate, liquids, culture mediums-Natural (Clots, Biological fluids, Tissue extracts), complex natural and chemically defined media
1.3. Explant-culture of explants, Cell culture technique- initiation, preparation and sterilization of media, isolation of explants, disaggregation of explants, culture, subculture
1.4.Cell lines, evolution of cell lines, maintenance of cell lines, Large scale culture of cell linesmonolayer, suspension and immobilized cell culture, Development of primary culture and cell lines, subculture
1.6. Stem cells: Characteristic features, maintenance, culture and Applications of Embryonic and adult stem cells, Animal cloning- Nuclear transfer and embryonic stem cell method

Unit-II Secondary Cultures
2.1. Cultured cells and evolution of continuous cell lines (established cell lines)
2.2. Commonly used cell lines – their origin and characteristics
2.3. Cell line preservation and characterization
2.4.High level expression of foreign gene in animal cells-expression vectors, enhancers, regulatory sequences. The need to express foreign genes in animal cells: advantage and disadvantages.
2.5. Molecular pharming: Transgenic animals and their applications, methods used for transgenesis with reference to transgenic mice ,cattle, sheep, goats, pigs, chicken and fish.

Unit-III Application of animal cell culture
3.1. Transfection methods of animal cells (Calcium phosphate, DEAE-dextran, Lipofection, Electroporation, Microinjection, Embryonic stem cell transfer)
3.2. Selection of recombinant cells with various marker genes (Thymidine Kinase, Dihydrofolate reductase, CAD protein, XGPRT, HAT, Neomycin phosphotransferase)
3.3. Production of transgenic animals (Mice, Cattle, Sheep, pigs, Fish and Birds)
3.4. Animal cells as a bioreactors for the production of commercially important products
3.5. Applications, advantages and disadvantages of animal tissue culture and Ethical issues related to transgenic animals.

REFERENCE BOOKS:

  1. Strategies in Transgenic Animal Sciences – By Glemn M.M. and James M. Robl ASM Press 2000.
  2. Practical Biotechnology – Methods and Protocols – By S. janarthanan and S. Vincent (Universities Press)
  3. Animal Cells as Bioreactors – By Terence Gartoright, Cambridge Univ Press
  4. Molecular Biotechnology – By Chinnarayappa (Universities Press)
  5. Principles and Practice of Animal Tissue Culture – By Sudha Gangal (Universities Press)
  6. Introduction to Veterinary Genetics – By F.W. Nicholas, Oxford University Press
  7. Biotechnology – By U. Satyanarayana
  8. Essentials of Biotechnology for Students – By Satya N. Das

ELECTIVE THEORY (B) ENVIRONMENTAL BIOTECHNOLOGY

Unit 1: Environmental Pollution
1.1 Introduction to environment and pollution
1.2 Types of pollution- air, water and land pollutions
1.3 Types of pollutants– inorganic, organic and biotic sources
1.4 Sources of pollution – domestic waste, agricultural waste, industrial effluents and municipal waste
1.5 Climate change, greenhouse gases and global warming, Impact of pollution on environment and measurement methods

Unit 2: Bioenergy and Bio-fuels
2.1 Renewable and non- renewable energy resources
2.2 Fossil fuels as energy source and their impact on environment
2.3 Non-conventional source – biomass as source of bioenergy
2.4 Types of biomass – plant, animal and microbial biomass
2.5 Production of biofuels, bioethanol & biomethanol

Unit 3: Bioremediation and Restoration of Environment
3.1 Microbial treatment of waste water (sewage of industrial effluent)- aerobic and anaerobic methods
3.2 Solid waste and management; Bioremediation– concepts and types (in-situ and ex-situ); Bioremediation of toxic metal ions– biosorption and bioaccumulation
3.3 Microbial bioremediation of pesticides and Xenobiotic compounds
3.4 Phytoremediation- concepts and application
3.5 Conservation of biodiversity

RECOMMENDED BOOKS:

  1. Text Book of Biotechnology – By H.K. Das (Wiley Publications)
  2. Biotechnology -By H.J. Rehm and G. Reed. VIH Publications, Germany
  3. Biogas Technology – By b.T. Nijaguna
  4. Biotechnology – By K. Trehan
  5. Industrial Microbiology – By L.E. Casida
  6. Food Microbiology – By M.R. Adams and M.O. Moss
  7. Introduction to Biotechnology – By P.K. Gupta
  8. Essentials of Biotechnology for Students – By Satya N. Das
  9. Bioprocess Engineering – By Shuler (Pearson Education)
  10. Essentials of Biotechnology – By Irfan Ali Khan and AtiyaKhanum (Ukaaz Publications)

GENERIC ELECTIVE BIOTECHNOLOGY PERSPECTIVES

Unit 1: Biotechnological applications in health care
1.1 Molecular diagnosis – monoclonal antibodies, DNA probes, Microarrays
1.2 DNA finger printing
1.3 Gene therapy
1.4 Recombinant therapeutic proteins – insulin, interferon, growth hormone
1.5 Stem cells and regenerative medicine
1.6 Transgenic animals – transgenic mice and transgenic fish

Unit 2: Biotechnological applications in agriculture and environment
2.1 Transgenic plants with insect resistance
2.2 Transgenic plants with drought stress tolerance
2.3 Bioremediation – Genetically engineered bacteria for bioremediation
2.4 Biofertilizers
2.5 Biopesticides
2.6 Biofuels

RECOMMENDED BOOKS:

  1. Human Molecular Genetics –Tom Strachen and A P Read, Bios Scxientific Publishers
  2. Human Genetics Molecular Evolution, Mc Conkey
  3. Recombinant DNA Technology, AEH Emery
  4. Principles and Practice of Medical Genetics, I, II, III Volumes by AEH Edts. Emery
  5. Medical Biotechnology-Pratibha Nallari, V.Venugopal Rao-Oxford Press
  6. Molecular Biotechnology-Glick
  7. Biodegradation and bioremediation Academic press By:San Diego
  8. Biotechnology in the sustainable environment, Plenumpress, NY
  9. Biotechnology- U.Satyanarayana
  10. Plant Tissue Culture By Akio Fujiwara
  11. Frontiers of Plant Tissue Culture By Trevor A. Thorpe
  12. Plant Tissue Culture: Theory and Practice By S.S. Bhojwani and A. Razdan