Literature DB >> 29091274

Screening Fusion Tags for Improved Recombinant Protein Expression in E. coli with the Expresso® Solubility and Expression Screening System.

Eric J Steinmetz1, Michele E Auldridge1.   

Abstract

The simplicity, speed, and low cost of bacterial culture make E. coli the system of choice for most initial trials of recombinant protein expression. However, many heterologous proteins are either poorly expressed in bacteria, or are produced as incorrectly folded, insoluble aggregates that lack the activity of the native protein. In many cases, fusion to a partner protein can allow for improved expression and/or solubility of a difficult target protein. Although several different fusion partners have gained favor, none are universally effective, and identifying the one that best improves soluble expression of a given target protein is an empirical process. This unit presents a strategy for parallel screening of fusion partners for enhanced expression or solubility. The Expresso® Solubility and Expression Screening System includes a panel of seven distinct fusion partners and utilizes an extremely simple cloning strategy to enable rapid screening and identification of the most effective fusion partner. © 2017 by John Wiley & Sons, Inc.
Copyright © 2017 John Wiley & Sons, Inc.

Entities:  

Keywords:  autoinduction; expresso; fusion tag; protein expression; solubility

Mesh:

Substances:

Year:  2017        PMID: 29091274      PMCID: PMC5791521          DOI: 10.1002/cpps.39

Source DB:  PubMed          Journal:  Curr Protoc Protein Sci        ISSN: 1934-3655


  24 in total

1.  Protein aggregation during overexpression limited by peptide extensions with large net negative charge.

Authors:  Yian-Biao Zhang; Jason Howitt; Sean McCorkle; Paul Lawrence; Karen Springer; Paul Freimuth
Journal:  Protein Expr Purif       Date:  2004-08       Impact factor: 1.650

Review 2.  Enhancement of soluble protein expression through the use of fusion tags.

Authors:  Dominic Esposito; Deb K Chatterjee
Journal:  Curr Opin Biotechnol       Date:  2006-06-15       Impact factor: 9.740

3.  Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency.

Authors:  R B Kapust; J Tözsér; J D Fox; D E Anderson; S Cherry; T D Copeland; D S Waugh
Journal:  Protein Eng       Date:  2001-12

4.  Release of proteins and peptides from fusion proteins using a recombinant plant virus proteinase.

Authors:  T D Parks; K K Leuther; E D Howard; S A Johnston; W G Dougherty
Journal:  Anal Biochem       Date:  1994-02-01       Impact factor: 3.365

5.  Rapid cloning by homologous recombination in vivo.

Authors:  P Bubeck; M Winkler; W Bautsch
Journal:  Nucleic Acids Res       Date:  1993-07-25       Impact factor: 16.971

6.  Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase.

Authors:  D B Smith; K S Johnson
Journal:  Gene       Date:  1988-07-15       Impact factor: 3.688

7.  Ubiquitin fusion augments the yield of cloned gene products in Escherichia coli.

Authors:  T R Butt; S Jonnalagadda; B P Monia; E J Sternberg; J A Marsh; J M Stadel; D J Ecker; S T Crooke
Journal:  Proc Natl Acad Sci U S A       Date:  1989-04       Impact factor: 11.205

8.  The thermo- and acido-stable ORF-99 from the archaeal virus AFV1.

Authors:  Adeline Goulet; Silvia Spinelli; Stéphanie Blangy; Herman van Tilbeurgh; Nicolas Leulliot; Tamara Basta; David Prangishvili; Christian Cambillau; Valérie Campanacci
Journal:  Protein Sci       Date:  2009-06       Impact factor: 6.725

9.  The ability to enhance the solubility of its fusion partners is an intrinsic property of maltose-binding protein but their folding is either spontaneous or chaperone-mediated.

Authors:  Sreejith Raran-Kurussi; David S Waugh
Journal:  PLoS One       Date:  2012-11-16       Impact factor: 3.240

10.  Optimization of an E. coli L-rhamnose-inducible expression vector: test of various genetic module combinations.

Authors:  Angelika Wegerer; Tianqi Sun; Josef Altenbuchner
Journal:  BMC Biotechnol       Date:  2008-01-14       Impact factor: 2.563

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  2 in total

1.  Selection of High Producers From Combinatorial Libraries for the Production of Recombinant Proteins in Escherichia coli and Vibrio natriegens.

Authors:  Joel Eichmann; Markus Oberpaul; Tobias Weidner; Doreen Gerlach; Peter Czermak
Journal:  Front Bioeng Biotechnol       Date:  2019-10-04

2.  Improving an Escherichia coli-based biocatalyst for terpenol glycosylation by variation of the expression system.

Authors:  Julian Rüdiger; Wilfried Schwab
Journal:  J Ind Microbiol Biotechnol       Date:  2019-05-06       Impact factor: 3.346

  2 in total

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