An improved system for expressing pancreatic ribonuclease in Escherichia coli

Abstract

An improved method for expressing and purifying bovine pancreatic ribonuclease from a synthetic gene using the λ promoter controlled by a temperature-sensitive repressor is described. The procedure involves isolation in the presence of a refolding buffer containing oxidized and reduced glutathione, under conditions where RNase can refold, but where proteases presumably do not. Yields are approx. 2 mg purified protein per 1 ferment