An Effective Recombinant Protein Expression and Purification System in Saccharomyces cerevisiae. Issue 1 (5th June 2018)

• Record Type: Journal Article
• Authors: Xie, Ying; Han, Xiao; Miao, Yansong
• Editors: Ausubel, Frederick M.; Brent, Roger; Kingston, Robert E.; Moore, David D.; Seidman, J.G.; Smith, John A.; Struhl, Kevin
• Is Part Of: Current protocols in molecular biology. Volume 123:Issue 1(2018)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2018-06-05
• Subjects: actin filament -- budding yeast -- protein purification -- post‐translational modification -- recombinant protein expression; 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/cpmb.62
• Languages: English
• ISSNs: 1934-3639

Abstract

Abstract: The expression and purification of recombinant proteins using bacterial vectors is a mature and preferred system to obtain folded and stable proteins. However, functional post‐translational protein modifications, such as glycosylation or phosphorylation, can only be achieved using eukaryotic expression systems. In addition, insolubility is another challenge when using proteins expressed in Escherichia coli, such as certain intrinsically disordered proteins, which are more prone to aggregation than folded proteins. Eukaryotic protein expression systems, including human cells, baculovirus/insect cells, and yeast, have become indispensable for the production of functional eukaryotic proteins. This article describes a detailed protocol for performing cytosolic protein expression, protein purification, and protein characterization using the budding yeast Saccharomyces cerevisiae . The introduced protein expression and purification system in yeast are advantageous due to the low cost, high yield, high protein solubility, and minimal expertise required. © 2018 by John Wiley & Sons, Inc.