An improved system for expressing pancreatic ribonuclease in Escherichia coli.
An improved method for expressing and purifying bovine pancreatic ribonuclease from a synthetic gene using the lambda 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.
McGeehan
G M
GM
Laboratory for Organic Chemistry, Swiss Federal Institute of Technology, Zurich.
Benner
S A
SA
D010582:Bacteriophage lambda / Q000235:genetics; D015153:Blotting, Western; D002384:Catalysis; D002850:Chromatography, Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005786:Gene Expression Regulation; D010179:Pancreas / Q000201:enzymology*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011489:Protein Denaturation; D012260:Ribonucleases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014169:Transformation, Bacterial
D012260:Ribonucleases