MICRB316 Lecture Notes - Lecture 5: Dna Footprinting, Intrinsically Disordered Proteins, Phosphodiester Bond

• School: University of Alberta
• Department: Microbiology (Biological Sciences)
• Course: MICRB316
• Professor: Nicolas Vozza

Document Summary

Rnap can recognize the upstream promoter element up. recognizes core promoter element (-10, -35); Gourse et al. mutations in a subunit led to some that made rnap incapable of responding to the up element. Wildtype rnap footprints both core and up element. A mutant rnap footprints core promoter only s. Therefore the a interaction with the up element. Footprinting is a method for detecting protein-dna interactions that can tell where the target site lies on the. A protein bound on a dna strand protects it from attack by something that will degrade the dna. Steps - dnase: end label dna (1 strand only) Dnase i under mild (low conc. ) conditions . 1 cut per strand strands, electrophorese beside untreated dna. Dms chemical is smaller and reveals more subtleties; Analysis shows a ntd and a ctd fold independently to form 2 domains tethered by a flexible linker. Clevage reveals that alpha domains are linked by 13 aa.