BPS 3101 Lecture Notes - Lecture 8: Dna-Binding Domain, Crystallography, Polyacrylamide Gel Electrophoresis

• School: University of Ottawa
• Department: Biopharmaceutical sciences
• Course: BPS 3101
• Professor: Linda Bonen

Document Summary

Start with gene of interest cloning, expression, purification of protein. X-ray crystallography: crystallize protein & x-ray diffraction analysis. Nmr spectroscopy: for small proteins or domains (in solution) Number of sequenced genomes: 2000 43, 2015 7211 (~200 fold) Identification of: protein motifs, catalytics centres, binding ligands, drugs, interaction with other macromolecules, relatedness to other proteins homology modelling i. e. zn finger, leu zipper (dna binding motifs in transcription factors) Clues from protein seq/ structure about biological function * Finding proteins that interact with protein of interest: phage coat protein & (cid:498)test protein(cid:499) from genome of interest: phage display: generate phage library producing collection of fusion proteins between. Hybrid protein will be displayed on outer surface of phage then you screen library to find ones having expressed protein which interacts with (cid:498)test(cid:499) protein: yeast 2-hybrid system: ( background info about transcription in eukaryotes) of interest. Transcription factors have 2 domains: transcription occurs (fit shapes) If tf domains physically separated no transcription.