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PEDS Advance Access published online on January 5, 2008
Protein Engineering Design and Selection, doi:10.1093/protein/gzm085
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© The Author 2007. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Short Communication

Engineering Escherichia coli heat-resistance by synthetic gene amplification

Daniel Christ1,2, Leo James, Jason W. Chin and Leo James1,3

1MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK

3 To whom correspondence should be addressed. E-mail: chin{at}mrc-lmb.cam.ac.uk

Organisms have evolved to exploit new environments by processes that involve both mutations and gene amplifications. Though in some cases amplified genes mutate to perform a different molecular function, in other cases altering gene copy number alone is sufficient to change organism function. Here we selected a library of genes, provided at high copy number, for their ability to confer survival on Escherichia coli cells at un-physiologically high temperatures. We find that a single gene (evgA), encoding a master transcriptional regulator, is overwhelmingly selected and allows survival upon heating to temperatures in excess of 50°C. While the detailed mechanisms of this resistance remained unclear, our results demonstrate the potential of copy number manipulation for the engineering of organisms

Keywords: copy number variation/synthetic biology/heat-resistance

Received August 28, 2007; revised October 22, 2007; accepted November 22, 2007.

2 Present address: Garvan Institute of Medical Research, Victoria Road, Darlinghurst, Sydney, NSW 2010, Australia


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