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Infection and Immunity, December 1998, p. 5599-5606, Vol. 66, No. 12
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Molecular and Functional Characterization of Salmonella enterica Serovar Typhimurium poxA Gene: Effect on Attenuation of Virulence and Protection

Koné Kaniga,1,* Melissa S. Compton,1 Roy Curtiss III,1,2 and Preeti Sundaram1

Megan Health, Inc., St. Louis, Missouri 63110,1 and Department of Biology, Washington University, St. Louis, Missouri 631302

Received 2 January 1998/Returned for modification 30 March 1998/Accepted 28 September 1998

Salmonella enterica poxA mutants exhibit a pleiotropic phenotype, including reduced pyruvate oxidase activity; reduced growth rate; and hypersensitivity to the herbicide sulfometuron methyl, alpha -ketobutyrate, and amino acid analogs. These mutants also failed to grow in the presence of the host antimicrobial peptide, protamine. In this study, PoxA- mutants of S. enterica serovar Typhimurium (S. typhimurium) were found to be 10,000-fold attenuated in orally inoculated BALB/c mice and 1,000-fold attenuated in intraperitoneally inoculated BALB/c mice, compared to wild-type S. typhimurium UK-1. In addition, poxA mutants were found to be capable of colonizing the spleen, mesenteric lymph nodes, and Peyer's patches; to induce strong humoral immune responses; and to protect mice against a lethal wild-type Salmonella challenge. A 2-kb DNA fragment was isolated from wild-type S. typhimurium UK-1 based on its ability to complement an isogenic poxA mutant. The nucleotide sequence of this DNA fragment revealed an open reading frame of 325 amino acids capable of encoding a polypeptide of 36.8 kDa that was confirmed in the bacteriophage T7 expression system. Comparison of the translated sequence to the available databases indicated high homology to a family of lysyl-tRNA synthetases. Our results indicate that a mutation of poxA has an attenuating effect on Salmonella virulence. Further, poxA mutants are immunogenic and could be useful in designing live vaccines with a variety of bacterial species. To our knowledge, this is the first report on the effect of poxA mutation on bacterial virulence.

* Corresponding author. Mailing address: Megan Health, Inc., 3655 Vista Ave., St. Louis, MO 63110. Phone: (314) 776-1626, ext. 104. Fax: (314) 776-3317. E-mail: kkaniga{at}meganhealth.com.

Infection and Immunity, December 1998, p. 5599-5606, Vol. 66, No. 12
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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