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Front Microbiol. 2014 Feb 25;5:67. doi: 10.3389/fmicb.2014.00067. eCollection 2014.

Generation of a vector suite for protein solubility screening.

Author information

1
Recombinant Protein Unit, Institut Pasteur de Montevideo Montevideo, Uruguay.
2
Protein Biophysics Unit, Institut Pasteur de Montevideo Montevideo, Uruguay.
3
Unité de Microbiologie Structurale, Institut Pasteur, Paris France.
4
Centre National de la Recherche Scientifique, Aix-Marseille Université CNRS UMR7257, AFMB, Marseille, France.

Abstract

Recombinant protein expression has become an invaluable tool for academic and biotechnological projects. With the use of high-throughput screening technologies for soluble protein production, uncountable target proteins have been produced in a soluble and homogeneous state enabling the realization of further studies. Evaluation of hundreds conditions requires the use of high-throughput cloning and screening methods. Here we describe a new versatile vector suite dedicated to the expression improvement of recombinant proteins (RP) with solubility problems. This vector suite allows the parallel cloning of the same PCR product into the 12 different expression vectors evaluating protein expression under different promoter strength, different fusion tags as well as different solubility enhancer proteins. Additionally, we propose the use of a new fusion protein which appears to be a useful solubility enhancer. Above all we propose in this work an economic and useful vector suite to fast track the solubility of different RP. We also propose a new solubility enhancer protein that can be included in the evaluation of the expression of RP that are insoluble in classical expression conditions.

KEYWORDS:

cloning; expression; high-throughput; recombinant proteins; solubility; vector

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