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Infection and Immunity, May 2005, p. 2818-2827, Vol. 73, No. 5
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.5.2818-2827.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Identification of Genes Required for Avian Escherichia coli Septicemia by Signature-Tagged Mutagenesis

Ganwu Li, Claudia Laturnus, Christa Ewers, and Lothar H. Wieler*

Institute of Microbiology and Epizootics, Free University Berlin, Philippstrasse 13, D-10115 Berlin, Germany

Received 13 September 2004/ Returned for modification 18 October 2004/ Accepted 3 January 2005

Infections with avian pathogenic Escherichia coli (APEC) cause colibacillosis, an acute and largely systemic disease resulting in significant economic losses in poultry industry worldwide. Although various virulence-associated genes have been identified in APEC, their actual role in pathogenesis is still not fully understood, and, furthermore, certain steps of the infection process have not been related to previously identified factors. Here we describe the application of a signature-tagged transposon mutagenesis (STM) approach to identify critical genes required for APEC infections in vivo. Twenty pools of about 1,800 IMT5155 (O2:H5) mutants were screened in an infection model using 5-week-old chickens, and potentially attenuated mutants were subjected to a secondary screen and in vivo competition assays to confirm their attenuation. A total of 28 genes required for E. coli septicemia in chickens were identified as candidates for further characterization. Among these disrupted genes, six encode proteins involved in biosynthesis of extracellular polysaccharides and lipopolysaccharides; two encode iron transporters that have not been previously characterized in APEC in in vivo studies, and four showed similarity to membrane or periplasmic proteins. In addition, several metabolic enzymes, putative proteins with unknown function, and open reading frames with no similarity to other database entries were identified. This genome-wide analysis has identified both novel and previously known factors potentially involved in pathogenesis of APEC infection.

* Corresponding author. Mailing address: Institute of Microbiology and Epizootics, Free University Berlin, P.O. Box 040225, Philippstraße 13, D-10115 Berlin, Germany. Phone: 49 030 2093 6300. Fax: 49 030 2093 6067. E-mail: wieler.lothar{at}vetmed.fu-berlin.de.

Editor: A. D. O'Brien

Infection and Immunity, May 2005, p. 2818-2827, Vol. 73, No. 5
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.5.2818-2827.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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