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Journal of Bacteriology, September 2003, p. 5301-5305, Vol. 185, No. 17
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.17.5301-5305.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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

James R. Johnson,1,2* Hank A. Lockman,3,{dagger} Krista Owens,1,2 Srdjan Jelacic,4,{ddagger} and Phillip I. Tarr4,§

Mucosal and Vaccine Research Center, Veterans Affairs Medical Center,1 Department of Medicine, University of Minnesota, Minneapolis, Minnesota 55417,2 Department of Pediatrics, Ohio State University, Columbus, Ohio 43210,3 Department of Pediatrics, University of Washington, Seattle, Washington 981954

Received 10 March 2003/ Accepted 20 June 2003

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.

* Corresponding author. Mailing address: VA Medical Center, Infectious Diseases (111F), One Veterans Dr., Minneapolis, MN 55417. Phone: (612) 467-4185. Fax: (612) 727-5995. E-mail: johns007{at}umn.edu.

{dagger} Present address: Battelle Memorial Institute, Columbus, OH 43201.

{ddagger} Present address: University of Washington School of Medicine, Seattle, WA 98195.

§ Present address: Department of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110.

Journal of Bacteriology, September 2003, p. 5301-5305, Vol. 185, No. 17
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.17.5301-5305.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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