Expression of bovine pancreatic ribonuclease A in Escherichia coli.
A synthetic gene for bovine pancreatic ribonuclease A (RNase A) has been expressed in Escherichia coli as a fusion protein with beta-galactosidase linked by the tetrapeptide Ile-Glu-Gly-Arg. RNase A was cleaved from the fusion using factor Xa, and the resulting product purified and reconstituted. The isolated RNase A was chromatographically, catalytically, and immunologically identical with authentic RNase A. This work argues that the method suggested by Nagai and Thogersen [Nagai, K. & Thogersen, H. C. (1984) Nature (Lond.) 309, 810-812] for releasing fusion proteins is quite general, even when applied to particularly complicated expression problem. The procedure here makes RNase A available for the first time as a model for studying structure-function relationships in proteins using site-directed mutagenesis.
D000818:Animals; D002384:Catalysis; D002417:Cattle; D004926:Escherichia coli / Q000235:genetics*; D005815:Genetic Code; D005822:Genetic Vectors; D010179:Pancreas / Q000201:enzymology*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D012259:Ribonuclease, Pancreatic / Q000235:genetics* / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D012259:Ribonuclease, Pancreatic