Article ID Journal Published Year Pages File Type
2020634 Protein Expression and Purification 2012 10 Pages PDF
Abstract

Producing recombinant proteins in Escherichia coli (E. coli) is generally performed using a trial and error approach with the different expression variables being tested independently from each other. As a consequence, variable interactions are lost which makes the trial and error approach quite time-consuming. In this paper, we report how switching from a trial and error to a fractional factorial approach allows testing in less than 2 weeks four expression variables (E. coli strains, culture media, expression temperatures and N-terminal fusion tags) in a single experiment. The method, called “Fusion-InFFact”, was validated using four test proteins. In all cases, Fusion-InFFact allowed finding conditions for expressing high yields of soluble proteins. The method was originally set-up for high throughput structural genomics programs, but can be used in any recombinant protein expression project.

► We describe a method for screening the expressing of recombinant proteins in Escherichia coli. ► The method is based on a fractional factorial approach combining five variables. ► We report the results obtained with four recombinant proteins.

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Life Sciences Biochemistry, Genetics and Molecular Biology Biochemistry
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