Embryonic stem cell differentiation: emergence of a new era in biology and medicine
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ANAT SC - Investigative Cell Biology | Course Outlines
Rates of enzymatic reactions. Enzymatic catalytic mechanisms: lysozyme and serine proteases.
Metabolism: Carbohydrate metabolism. Electron transport and oxidative phosphorylation. Photosynthesis Lipid metabolism.
Amino acid metabolism. Nucleotide metabolism. Integration of metabolic pathways and metabolic disorders. Recommended Reading D. Voet and J. Voet, Biochemistry , 4th Edn. Nelson and M. Cox, Lehninger Principles of Biochemistry , 5th Edn. Beginning the Developmental Program: Gametogenesis, Structure of eggs and sperm, Comparing oogenesis and spermatogenesis 2. Segment identity and Hox genes. Models of Cell Differentiation Introduction to developmental genetics techniques and concepts Completing the Life Cycle, and New Beginnings— Sex determination, Dosage compensation, Imprinting Introduction to concepts in stem cell biology renewal, potency, etc.
Development and Evolution: Developmental mechanisms of evolutionary change Evolution of morphological novelty Recommended Reading S. Gilbert, Developmental Biology , 8th Edn. Wolpert, J. Smith, T. Jessell, P. Lawrence, E. Robertson and E. Meyerowitz, Principles of Development , 3rd Edn. Respiration in plants: Respiration during photosynthesis, role of environmental factors on rate of respiration Regulation of Plant growth: Gibberlins, Auxins, Cytokines, Ethylene, Abscisic acid, Brassinosteroids Reproduction in flowering plants: transition from vegetative phase to flowering state.
Plant protection: Plant defence against pathogens, insects and herbivores. Plant-plant interaction: Allelopathy Recommended Reading N. Taiz and E. Zeiger, Plant Physiology , 4th Edn. Analysis of DNA sequence for presence of internal restriction digestion sites etc. PCR amplification of the DNA segment of interest from a suitable source, purification of the PCR product, restriction digestion, and subsequent ligation into the suitable bacterial expression vector also containing an antibiotic resistant marker of interest.
Effect of substrate stiffness on early human embryonic stem cell differentiation
Transformation into suitable competent cells BL21 etc. Students will learn how to prepare E. Selection of the antibiotic resistant single colony.
Plasmid isolation from the transformed cells and sequencing it to confirm the sequence of cloned DNA segment of interest. Large scale expression and purification of the protein: Bulk scale bacterial cell culture and IPTG induction for protein expression. Detection of the protein by western blotting in soluble and insoluble fraction after bacterial cell lysis.
KANEKO Complex Systems Biology
Affinity purification of the protein from the soluble fraction of the bacterial cell lysate for His-tagged protein, Ni-agarose matrix will be used. Biochemical and biophysical characterizations of the purified protein: Purified protein will be assayed for its biological activity. Fluorescence from GFP. Fluorescence quenching and polarization studies. Unfolding and refolding studies using CD and fluorescence methods.
Sambrook, E. Fritsch, and T. Maniatis, Molecular cloning: a laboratory manual , 2nd Edn. The choice of model organisms to be used for these experiments is up to the discretion of the instructor. An outline of some of the aspects that can be explored is as follows: Cleavage patterns. Segmentation during embryonic development; Reporter gene expression; mutant phenotypes. Master regulators in cell fate specifications. Pattern formation in developing tissues Programmed. Cell Death and tissue sculpturing Role of niche and stem cells in tissue regeneration.
Generation of action potential; neuronal propagation and synaptic transmission. Nucleosomes, histones, chromatids, centromeres, teleomeres. Repetitive content of eukaryotic genomes.
- Cell division.
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Organization of prokaryotic genomes. Organization of genes, operons. Mapping of Genomes.