ARS/NET Agricultural Biotechnology Syllabus

Unit 1: Cell Structure and Function

Prokaryotic and eukaryotic cell architecture. Cell wall, plasma membrane. Structure and function of cell organelles: vacuoles, mitochondria, plastids, Golgi apparatus, ER, peroxisomes, glyoxisomes. Cell division, regulation of cell cycle. Protein secretion and targeting. Cell division, growth and differentiation.

Unit 2: Biomolecules and Metabolism

Structure and function of carbohydrates, lipids, proteins, and nucleic acids. Synthesis of carbohydrate, glycolysis, HMP, Citric acid cycle and metabolic regulation. Oxidative phosphorylation and substrate level phosphorylation. Vitamins, plant hormones. Functional molecules, antioxidants, nutrient precursors, HSPs, anti-viral compounds.

Unit 3: Enzymology

Enzymes: structure, conformation, classification. Assay, isolation, purification, and characterization. Catalytic specificity, mechanism of action, active site. Regulation of enzyme activity.

Unit 4: Molecular Genetics

Concept of gene. Prokaryotes as genetic system. Prokaryotic and eukaryotic chromosomes. Methods of gene isolation and identification. Split genes, overlapping genes, and pseudogenes. Organization of prokaryotic and eukaryotic genes and genomes including operon, exon, intron, enhancer, promoter sequences, and regulatory elements. Mutation: spontaneous, induced, and site-directed. Recombination in bacteria, fungi, and viruses. Transformation, transduction, conjugation. Transposable elements and transposition.

Unit 5: Gene Expression

Expression of genetic information. Operon concept. Transcription mechanisms in prokaryotes and eukaryotes. Transcription unit, regulatory sequences, enhancers, activators, repressors, co-activators, co-repressors. Inducible genes and promoters. Transcription factors, post-transcriptional modification, and protein transport. DNA-protein interaction. Genetic code. Mechanism of translation and its control. Post-translational modifications.

Unit 6: Molecular Biology Techniques

Isolation and purification of nucleic acids. Nucleic acids hybridization: Southern, Northern, and Western blotting. Immune response, monoclonal and polyclonal antibodies, ELISA. DNA sequencing. Construction and screening of genomic and cDNA libraries. Gel electrophoretic techniques. PCR, RT-PCR, qRT-PCR. Spectroscopy, ultracentrifugation, chromatography. FISH, RIA, etc.

Unit 7: Gene Cloning

Restriction enzymes and their uses. Features and uses of vectors: plasmids, bacteriophages, phagmids, cosmids, BACs, PACs, YACs, binary vectors, expression vectors. Gene cloning and sub-cloning strategies. Chromosome walking. Genetic transformation. Risk assessment and intellectual property rights (IPR).

Unit 8: Molecular Biology

Ribosome structure and function. Protein biosynthesis in prokaryotes and eukaryotes. Post-translational modification. Gene regulation. RNA processing and post-transcriptional modifications. Bioprospecting, biofortification, gene pyramiding and gene fusion. Ribozyme technology.

Unit 9: Plant Molecular Biology

Photoregulation and phytochrome regulation of nuclear and chloroplastic gene expression. Photosynthesis: C3 & C4 cycle and photorespiration. Molecular mechanism of nitrogen fixation. Molecular biology of abiotic stresses (drought, salt, heavy metals, temperature). Molecular biology of biotic stresses (bacterial, fungal, viral diseases). Signal transduction and molecular basis. Molecular mechanism of plant hormone action. Mitochondrial control of fertility. Structure, organization, and regulation of nuclear genes related to storage proteins and starch synthesis.

Unit 10: Tissue Culture

Basic techniques in cell culture and somatic cell genetics. Clonal propagation. Concept of cellular totipotency. Anther culture, somaclonal and gametoclonal variations. Hybrid embryo culture and embryo rescue. Somatic hybridization and cybridization. Application of tissue culture in crop improvement. Secondary metabolite production. In vitro mutagenesis, cryopreservation, and plant tissue culture repository.

Unit 11: Plant Genetic Engineering

Isolation of genes of economic importance. Gene constructs for tissue-specific expression. Methods of gene transfer to plants: direct and vector-mediated. Molecular analysis of transformants. Applications in crop improvement: insect-pest resistance, abiotic stress resistance, herbicide resistance, storage protein quality, increasing shelf-life, oil quality. Current status of transgenics, biosafety norms, controlled field trials, and release of transgenics (GMOs).

Unit 12: Molecular Markers and Genomics

DNA molecular markers: principles, types, and applications. RFLP, AFLP, RAPD, SSR, SNP. Structural and functional genomics. Gene mapping, genome mapping, gene tagging. Transcriptomics, ionomics, metabolomics, and proteomics. Comparative genomics and applications.

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