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Genomics II: The Proteome

Genomics II: The Proteome. Using high-throughput methods to identify proteins and to understand their function. Protein Complementation. Enzymatic complementation b -galactosidase reconstitution Fluorescence complementation GFP or YFP reconstitution

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Genomics II: The Proteome

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  1. Genomics II:The Proteome Using high-throughput methods to identify proteins and to understand their function

  2. Protein Complementation • Enzymatic complementation • b-galactosidase reconstitution • Fluorescence complementation • GFP or YFP reconstitution • FRET (fluorescence resonance energy transfer)

  3. Enzymatic Complementation

  4. Blue=DAPI Red=BGAL Blue=DAPI Green=BGAL Red=Actin

  5. Bimolecular Fluorescence Complementation

  6. FRET (fluorescence resonance energy transfer)

  7. Summary I • Goals of proteomics • Identify and ascribe function to proteins under all biologically plausible conditions • Proteomics methods • 2-D gel electrophoresis for separating proteins on the basis of charge and molecular weight • Mass spectrometry for identifying proteins by measuring the mass-to-charge ratio of their ionized peptide fragments • Protein chips to identify proteins, to detect protein–protein interactions, to perform biochemical assays, and to study drug–target interactions

  8. Summary II • Proteomics methods (continued) • Yeast two-hybrid method for studying protein–protein interactions • Biochemical genomics for high-throughput assays • Some accomplishments of proteomics • Example: yeast • Yeast two-hybrid method reveals interactome • Transcriptional regulatory networks deduced • Biochemical genomics uncovers new ORF functions • Subcellular localization of proteins

  9. Linkage and Mapping

  10. Linked alleles tend to be inherited together

  11. Crossing over produces new allelic combinations

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