PEDS Advance Access originally published online on January 5, 2008
Protein Engineering Design and Selection 2008 21(2):121-125; doi:10.1093/protein/gzm085
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Short communication |
Engineering Escherichia coli heat-resistance by synthetic gene amplification
1MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK
3 To whom correspondence should be addressed. E-mail: d.christ{at}garvan.org.au (D.C.); E-mail: chin{at}mrc-lmb.cam.ac.uk (J.W.C.)
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.