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Cullen Lecture 4: Genomics and Proteomics (A Techniques-Based Lecture) pjcullen@buffalo

Cullen Lecture 4: Genomics and Proteomics (A Techniques-Based Lecture) pjcullen@buffalo.edu. DNA Microarray Analysis Two-hybrid Analysis Synthetic Genetic Analysis. DNA Microarrays. A separate PCR product for each gene in the genome.

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Cullen Lecture 4: Genomics and Proteomics (A Techniques-Based Lecture) pjcullen@buffalo

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  1. Cullen Lecture 4: Genomics and Proteomics (A Techniques-Based Lecture) pjcullen@buffalo.edu

  2. DNA Microarray Analysis Two-hybrid Analysis Synthetic Genetic Analysis

  3. DNA Microarrays A separate PCR product for each gene in the genome Every gene is represented by a unique spot of DNA on the slide

  4. DNA Microarrays The microarray can then be used to compare the expression of all of the genes in the genome simultaneously

  5. DNA Microarrays After hybridization, a scanner is used to measure the fluorescence signal from each spot Green -- expression of the gene is higher after drug treatment Red -- expression of the gene is lower after drug treatment Yellow (Red:Green = 1) -- no change in the expression of the gene

  6. DNA Microarrays Data from multiple microarray experiment can be grouped and analyzed to identify genes that are coordinately regulated. E.g. Part of the transcriptional pattern of genes in a fibroblast responding to serum stimulation:

  7. Targets of the FG pathway

  8. DNA Microarrays Use as a diagnostic tool

  9. DNA Microarrays Use as diagnostic tool Green spots identify genes upregulated in the tumor Red spots genes that are downregulated in the tumor

  10. Genome-Wide Genetic Screens MSB2 was identified in both screens

  11. Synthetic Genetic Analysis Gene 1 (mutant 1) Gene 2 (mutant 2) Process 2 Process 1 Process 3 Shared Function (dead)

  12. SGA analysis on a Genome-wide Scale Mutant 1 + Mutant 2 (all deletions) pGENE1 FOA (Mutant 1 and Mutant 2) Lacking pGENE1 (dead?) MSB2 was identified in both screens

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