High-Frequency Secondary Mutations after Suicide-Driven Allelic Exchange Mutagenesis in Extraintestinal Pathogenic Escherichia coli

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Abstract
Frequent unintended secondary mutations occurred in extraintestinal pathogenic Escherichia coli strains CP9, CFT073, and RS218 during suicide plasmid-mediated, putatively specific deletions of hlyA , papG allele III, and iha . Pulsed-field gel electrophoresis and PCR analyses demonstrated genomic alterations and/or unintended loss of defined virulence genes ( papG , the F7-2 papA allele, iutA , sat , hlyD , and cnf ). Caution is warranted when attributing the observed phenotypic changes to the intended mutation.

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