lecture18

Is a particular tf required what genes are cell type

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Unformatted text preview: articular TF required? What genes are cell type-specific? Are there gene expression “signatures” that indicate a disease? 1817 Chromatin Immunoprecipitation: ChIP Crosslink protein to DNA in living cells with formaldehyde Break open cells/ Add primary antibody shear DNA of interest Add Protein A agarose beads Input IP’ed Input IP’ed PCR amplify (Look for enrichment of precipitated DNA) Reverse crosslinks/ remove proteins IP to enrich for DNA bound by protein of interest Adapted from Active Motif® scheme www.activemotif.com/downloads/profile_pdfs/ chip-it_profile.pdf 1818 ChIP-chip 1819 Determining the in vivo binding site specificity of TFs 1820 Mapping all TF binding sites for all genes 1821 ChIP-chip: some applications Determining the in vivo binding site specificity of TFs Is it the same to in vitro data? If not, why not? Mapping all binding sites for a particular TF Are target genes different in different cell types ? How about healthy vs. diseased c...
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This note was uploaded on 10/30/2010 for the course CHEM 114C taught by Professor Alexanderhoffmann during the Spring '08 term at UCSD.

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