Biochemistry 3382A Study Guide - Final Guide: Short Hairpin Rna, Orf 2, Rna Interference

• School: Western University
• Department: Biochemistry
• Course: Biochemistry 3382A
• Professor: David Litchfield

Document Summary

Rip-chip: mild lysis of cell->ip using antibodies against protein of interest- Par-clip: cell treated with photoactivatable nucleosides->cross linking by activation with uv 365 nm->mild lysis->ip->unbound rna around protein digest- >rna of intresest labeled->sds page, elution->protein removed by proteinase k- >convert to cdna->pcr->sequence [par clip can also be used to cross link and indentify microrna-mrna interactions due to the photoactivatable nucleosides] (clip similar but doesn"t use photoactivable nucleoside, less specific cross linking and less efficient) Tran acting elements- aubps, they can promote degradation by exosome by destabilizing the element. Loss of the ability for trans-acting elements to bind this region causes a loss of control over the transcript expression in response to external stimuli and plays a role in cancer. Amd also plays a role in the regulation of genes during an immune response. Exosome:************** definite exam question multi-enzyme complex of 9 essential proteins including the exoribonuclease rrp44p.