کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2020634 | 1069195 | 2012 | 10 صفحه PDF | دانلود رایگان |

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.
Journal: Protein Expression and Purification - Volume 84, Issue 2, August 2012, Pages 204–213