Expression of bovine pancreatic ribonuclease A in Escherichia coli
- 1 February 1987
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 163 (1) , 67-71
- https://doi.org/10.1111/j.1432-1033.1987.tb10737.x
Abstract
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. and Thogersen, H. C. (1984) Nature (Lond.) 309, 810-812] for releasing fusion proteins is quite general, even when applied to a 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.This publication has 39 references indexed in Scilit:
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