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Infection and Immunity, March 2003, p. 1343-1351, Vol. 71, No. 3
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.3.1343-1351.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Functional Genomics Approach to the Identification of Virulence Genes Involved in Edwardsiella tarda Pathogenesis

Putanae S. Srinivasa Rao,1 Tit Meng Lim,1 and Ka Yin Leung1,2*

Department of Biological Sciences, Faculty of Science,1 Tropical Marine Science Institute, National University of Singapore, Singapore 117543, Singapore2

Received 22 July 2002/ Returned for modification 13 November 2002/ Accepted 9 December 2002

Edwardsiella tarda is an important cause of hemorrhagic septicemia in fish and also of gastro- and extraintestinal infections in humans. Here, we report the identification of 14 virulence genes of pathogenic E. tarda that are essential for disseminated infection, via a genome-wide analysis. We screened 490 alkaline phosphatase fusion mutants from a library of 450,000 TnphoA transconjugants derived from strain PPD130/91, using fish as an infection model. Compared to the wild type, 15 mutants showed significant decreases in virulence. Six mutants had insertions in the known virulence-related genes, namely, fimA, gadB, katB, pstS, pstC, and ssrB. Some mutants corresponded to known genes (astA, isor, and ompS2) that had not been previously shown to be involved in pathogenesis, and three had insertions in two novel genes. In vivo infection kinetics experiments confirmed the inability of these attenuated mutants to proliferate and cause fatal infection in fish. Screening for the presence of the above-described virulence genes in six virulent and seven avirulent strains of E. tarda indicated that seven of the genes were specific to pathogenic E. tarda. The genes identified here may be used to develop vaccines and diagnostic kits as well as for further studying the pathogenesis of E. tarda and other pathogenic bacteria.

* Corresponding author. Mailing address: Department of Biological Sciences, Faculty of Science, National University of Singapore. Science Dr. 4, Singapore 117543, Singapore. Phone: (65) 6874 7835. Fax: (65) 6779 2486. E-mail: dbslky{at}nus.edu.sg.

Editor: B. B. Finlay

Infection and Immunity, March 2003, p. 1343-1351, Vol. 71, No. 3
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.3.1343-1351.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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