Recombinant Expression, Isotope Labeling, and Purification of Cold Shock Protein from Colwellia psychrerythraea for NMR Study

Abstract

Cold shock proteins (Csps) are a subgroup of the cold-induced(I proteins oil reduction of file growth temperature below the physiological temperature They preferentially bind to single-stranded nucleic acids to translational regulation via RNA chaperoning. Csp plays Important role in cold adaptations for the psychrophilic microorganism Recently, Cold shock protein From psychrophilic bacteria, Colwellia psychrerythraea (CpCsp) has been identified Three dimensional Structures of a number of Csps from various microorganisms have been solved by NMR spectroscopy or X-ray crystallography, but Structures of psychrophilic Csps were not studied yet Therefore, cloning and purification protocols for further structural study of psychrophilic Csp have been optimized in tills study CpCsp was expressed in E coli with pET-11a vector system and purified by ion exchange, size exclusion, and reverse phase chromatography. Expression and purification of CpCsp in M9 minimal media was carried out and (15)N-labeled proteins with high purity over 90% was obtained. Further Study will be carried Out to investigate the tertiary Structure and dynamics of CpCsp.