B.Sc. (Hons) Biochemistry Syllabus

Semester VI

  • COURSE CODE
    COURSE NAME
    CREDITS
  • JBBC-601

    Metabolisms of Amino acids & Nucleotides:

    UNIT-I

    Outlines of Amino Acids metabolism: Digestion, absorption and uptake of Amino Acids including γ-glutamyl cycle; Transamination, role of PLP, oxidative and nonoxidative deamination, glucosealanine cycle, urea cycle and inherited defects of urea cycle, Krebs's bicycle.

    UNIT-II

    Degradation of the carbon skeleton: Glucogenic and ketogenic amino acid catabolic pathways for the 20 standard amino acids; Metabolism of one-carbon units.

    UNIT-III

    Disorders of amino acid metabolism: Phenylketonuria, Alkaptonuria, Maple syrupurine disease, Methylmalonic aciduria, Parkinson's disease, Homocystinuria, and Hartnup's disease

    UNIT-IV

    Biosynthesis of purine nucleotides: Biosynthesis of IMP; pathways from IMP to AMP and GMP; conversion to triphosphates; regulation of purine Nucleotide biosynthesis, salvage pathways; synthesis of coenzymes (NAD+, FMN, FAD, HSCoA) Biosynthesis of pyrimidine nucleotides: Biosynthesis of UMP, conversion of triphosphate and regulation of Biosynthesis of pyrimidine nucleotide synthesis.

    UNIT-V

    Degradation of purine and pyrimidine nucleotides. Disorders of nucleotide metabolism: Lesch Nyhan syndrome, Gout, SCID.

    Books:

    • 1. Cox, M.M. and Nelson, D.L. (2008). Lehninger Principles of Biochemistry, W.H. Freeman and Company, New York, USA
    • 2. Voet, D. and Voet, J.G. (2004). Biochemistry, John Wiley and Sons. INC.
    • 3. Devlin, T.M. (2002) Textbook of Biochemistry with clinical correlations, John Wiley and sons, INC.
    04
  • JBBC-602

    Biochemical Techniques:

    UNIT-I

    Separation Techniques: Different methods of protein precipitation: Precipitation using inorganic salts (salting out) and organic solvents, isoelectric precipitation, Dialysis, Ultrafilteration, Lyophilization.

    UNIT-II

    Chromatography: Basic principles of chromatography: Partition coefficient, concept of theoretical plates, various modes of chromatography (paper, thin layer, column), preparative and analytical applications, LPLC and HPLC. Different types of chromatography: Paper Chromatography, Thin Layer Chromatography. Molecular Sieve Chromatography, Ion Exchange Chromatography, Affinity Chromatography, Gas Liquid Chromatography

    UNIT-III

    Electrophoresis: Basic Principle of electrophoresis, Paper electrophoresis, Gel electrophoresis, discontinuous gel electrophoresis, PAGE, SDS-PAGE, Native gels, denaturing gels, agarose gel electrophoresis, buffer systems in electrophoresis, electrophoresis of proteins and nucleic acids, protein and nucleic acid blotting, detection and identification (staining procedures), molecular weight determination, Isoelectric Focusing of proteins.

    UNIT-IV

    Principle of centrifugation, basic rules of sedimentation, sedimentation coefficient, various types of centrifuges, different types of rotors, differential centrifugation, density gradient centrifugation (Rate zonal and Isopycnic).

    UNIT-V

    Spectrophotometry: Principleof UV-Visible absorption spectrophotometry, instrumentation and applications

    Books:

    • 1. Freifelder, D., Physical Biochemistry, W.H. Freeman and Co., N.Y. USA.
    • 2. Cooper, T.G., The Tools of Biochemistry John Wiley and Sons, N.Y. USA.
    04
  • JBBC-603

    Recombinant DNA Technology:

    UNIT-I

    Concept and emergence of r-DNA technology: Basic techniques involved in rDNA technology. Restriction Enzymes, DNA methylation systems in E.coli, other enzymes used in cloning (DNA polymerases, RNA Polymerases, Reverse Transcriptase, Ligases, Taq polymeraseetc.) Cloning vectors – Plasmids, λ bacteriophage based, M13 phage based, phagemids. High capacity vectors: Cosmids, yeast artificial chromosomes, bacterial artificial chromosomes, Covalent linkage of DNA fragments to vector molecules: Linkers, Adapters, homopolymer tailing. Generation of genomic and cDNA libraries (different methods of cDNA synthesis), Solid phase synthesis of DNA.

    UNIT-II

    Nucleic acid blotting: Southern, Northern, Western, dot and slot blot. Selection and screening of recombinant clones: Probe preparation (radiolabelled and non radiolabelled) via nick translation, random priming etc. Guessmers and degenerate probes. Sequence dependent and independent screening, southern-western, colony and plaque hybridization, in situ chromosomal hybridization, chromosome walking, etc.).

    UNIT-III

    Expression of cloned DNA: Expression vectors (lac promoter, tryptophan promoter, Lambda cI promoter, arabinose promoter based) optimization of protein expression in heterologous systems (using upstream and downstream signals) Fusion proteins, In vitro translation systems. RNAi vectors.

    UNIT-IV

    DNA transactions in Microorganisms: Cloning DNA/RNA in bacteria (Transformation, transduction and conjugation), methods of gene transfer into yeast (YIp, YEp, YCp, YRp, shuttle vectors) fungi, plant and animal host systems. Characterization of cloned DNA: Restriction mapping. DNA sequencing (dideoxychain termination, chemical degradation, shotgun sequencing, contig assembly and pyrosequencing etc.) Polymerase Chain Reaction, VNTRs, DNA fingerprinting, SNPs, RFLPs. Modification of cloned DNA: Site directed mutagenesis, Protein engineering.

    UNIT-V

    Applications of recombinant DNA technology: Production of recombinant proteins in bacterial and eukaryotic cells – Recombinant insulin, growth hormone, factor VIII, recombinant vaccines etc. Identification of genes responsible for human diseases, diagnostics, gene therapy. Genetically modified plants. Ethical, legal and social issues.

    Books:

    • 1. Glick, B.R. and Pasternak, J.J., Molecular Biotechnology: Principles and applications of recombinant DNA technology. Asm Press, Washington, USA.
    • 2. Primrose, S.B. and Twyman. R.M. Principles of gene manipulation and genomics. Blackwell Publishing, MA, USA.
    • 3. Sambrook, J. and Russell, D., Molecular cloning: a laboratory manual, 3rd Edition. Cold Spring Harbor Laboratory Press, New York.
    04
  • JBBC-651

    Metabolisms of Amino acids & Nucleotides Practical:

    Based on theory paper of Metabolisms of Amino acids & Nucleotides.
    02
  • JBBC-652

    Biochemical Techniques Practical:

    Based on theory paper of Biochemical Techniques.
    02
  • JBBC-653

    Recombinant DNA Technology Practical:

    Based on theory paper of Recombinant DNA Technology.
    02
  • Total Credits
     
    18