Production of Disulfide‐Bonded Proteins in Escherichia coli. Issue 1 (1st October 2014)

• Record Type: Journal Article
• Authors: Ke, Na; Berkmen, Mehmet
• Is Part Of: Current protocols in molecular biology. Volume 108:Issue 1(2014)
• Page Start: 16.1B.1
• Page End: 16.1B.21
• Publication Date: 2014-10-01
• Subjects: disulfide‐bonded protein production -- E. coli -- SHuffle; Molecular biology -- Technique -- Periodicals; Molecular biology -- Laboratory manuals; Molecular Biology -- methods; Biologie moléculaire -- Technique; Biologie moléculaire -- Manuels de laboratoire; Molecular biology; Molecular biology -- Technique; Laboratory Manual; Electronic reference sources; Laboratory manuals; 572.8028
• DOI: 10.1002/0471142727.mb1601bs108
• Languages: English
• ISSNs: 1934-3639

Abstract

Abstract: Production of recombinant proteins at high yields in Escherichia coli requires extensive optimization of expression conditions. Production is further complicated for proteins that require specific post‐translational modifications for their eventual folding. One common and particularly important post‐translational modification is oxidation of the correct pair of cysteines to form a disulfide bond. This unit describes methods to produce disulfide‐bonded proteins in E. coli in either the naturally oxidizing periplasm or the cytoplasm of appropriately engineered cells. The focus is on variables key to improving the oxidative folding of disulfide‐bonded proteins, with the aim of helping the researcher optimize expression conditions for a protein of interest. Curr. Protoc. Mol. Biol . 108:16.1B.1‐16.1B.21. © 2014 by John Wiley & Sons, Inc.