PCL366H1 Study Guide - Final Guide: Striatum, Sphericity, Observational Error
23 Sep 2016
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Matthews chromatin immunoprecipitation assay (chip) and chip-chip/seq to study transcription factor function. His research focus is on the aryl hydrocarbon receptor and estrogen receptor: ahr is a cytosolic nr bound to chaperone proteins, upon ligand binding, it dissociates from chaperones and enters nucleus, dimerizing with. It binds ahre, dre, or xre (response elements) and recruits coactivators upregulats. Recognized sequence: tngngtg: lollipop box, estrogen receptor (from pcl302) It aims to determine whether specific proteins are associated with specific genomic regions, such as transcription factors on promoters(cid:863) wikipedia. Establish protein interactions with in vivo dna sequences. Allows study of histone modifications on transcription in vivo. Methodology: treating cells or tissues briefly with formaldehyde. Formaldehyde crosslinks dna and protein (cytosine, lysine, arginine). Cell nuclei are isolated (though whole cell extracts work too: fragmenting the dna/protein complexes. More sonication = less smearing of bands. Samples are centrifuged and supernatant (chromatin) is isolated. Add antibodies specific to protein of interest.
