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Open AccessCommunication

Application and Optimization of relE as a Negative Selection Marker for Making Definitive Genetic Constructs in Uropathogenic Escherichia coli

1
Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, 1E Kent Ridge Road, NUHS Tower Block, Level 10, Singapore 119074, Singapore
2
Genome Institute of Singapore, Infectious Diseases Group, 60 Biopolis street, Genome, #02-01, Singapore 138672, Singapore
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Catharina Svanborg
Pathogens 2016, 5(1), 9; https://doi.org/10.3390/pathogens5010009
Received: 11 September 2015 / Revised: 7 January 2016 / Accepted: 13 January 2016 / Published: 18 January 2016
(This article belongs to the Special Issue Molecular Aspects of Urinary Tract Infection)
Studies of Uropathogenic Escherichia coli (UPEC) pathogenesis have relied heavily on genetic manipulation to understand virulence factors. We applied a recently reported positive-negative selection system to create a series of unmarked, scarless FimH mutants that show identical phenotypes to previously reported marked FimH mutants; these are now improved versions useful for definitive assignment of phenotypes to FimH mutations. We also increased the efficiency of this system by designing new primer sites, which should further improve the efficiency and convenience of using negative selection in UTI89, other UPEC, and other Enterobacteriaceae. View Full-Text
Keywords: negative selection; UTI89; FimH negative selection; UTI89; FimH
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Khetrapal, V.; Mehershahi, K.S.; Chen, S.; Chen, S.L. Application and Optimization of relE as a Negative Selection Marker for Making Definitive Genetic Constructs in Uropathogenic Escherichia coli. Pathogens 2016, 5, 9.

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