Tandem Cell‐Free Protein Synthesis as a Tool for Rapid Screening of Optimal Molecular Chaperones. Issue 5 (15th April 2019)

• Record Type: Journal Article
• Authors: Yang, Hyeon‐Jung; Lee, Kyung‐Ho; Lim, Hye Jin; Kim, Dong‐Myung
• Is Part Of: Biotechnology journal. Volume 14:Issue 5(2019)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2019-04-15
• Subjects: cell‐free protein synthesis -- molecular chaperone -- screening -- tandem expression -- transient cell‐free expression; Biotechnology -- Periodicals; 660.605
• DOI: 10.1002/biot.201800523
• Languages: English
• ISSNs: 1860-6768

Abstract

Abstract : A simple and flexible method is developed for rapid screening of molecular chaperones that enhance the functional expression of recombinant proteins. A panel of molecular chaperones are transiently expressed in a reaction mixture of cell‐free protein synthesis and then a target protein is subsequently expressed in the presence of these presynthesized molecular chaperones. The biological activity of the cell‐free synthesized target protein is compared to identify the effective molecular chaperones. This strategy successfully identifies individual and combinations of bacterial molecular chaperones that markedly improved the functional expression of horseradish peroxidase. The authors believe that the presented strategy provides a versatile platform for the optimal production of functional proteins, and can also be extended to studies of other interacting proteins. Abstract : A method for rapid screening of molecular chaperones is described. In this method, a series of molecular chaperones and a target protein are sequentially synthesized in a cell‐free synthesis system to determine the effect of the priorly synthesized molecular chaperones on the folding of the target protein. This method allows rapid selection of molecular chaperones that are optimal for the functional synthesis of recombinant proteins.