This Article Full Text Full Text (PDF) Supplemental material Other Versions of this Article: JB.01735-07v1 190/7/2306    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Schell, M. A. Articles by DeShazer, D. Search for Related Content PubMed PubMed Citation Articles by Schell, M. A. Articles by DeShazer, D.

 Previous Article  |  Next Article 

Journal of Bacteriology, April 2008, p. 2306-2313, Vol. 190, No. 7
0021-9193/08/$08.00+0     doi:10.1128/JB.01735-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Comparative Genomics and an Insect Model Rapidly Identify Novel Virulence Genes of Burkholderia mallei ^,

Department of Microbiology, University of Georgia, Athens, Georgia 30602,¹ Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland 21702-5011²

Received 30 October 2007/ Accepted 8 January 2008

Burkholderia pseudomallei and its host-adapted deletion clone Burkholderia mallei cause the potentially fatal human diseases melioidosis and glanders, respectively. The antibiotic resistance profile and ability to infect via aerosol of these organisms and the absence of protective vaccines have led to their classification as major biothreats and select agents. Although documented infections by these bacteria date back over 100 years, relatively little is known about their virulence and pathogenicity mechanisms. We used in silico genomic subtraction to generate their virulome, a set of 650 putative virulence-related genes shared by B. pseudomallei and B. mallei but not present in five closely related nonpathogenic Burkholderia species. Although most of these genes are clustered in putative operons, the number of targets for mutant construction and verification of reduced virulence in animal models is formidable. Therefore, Galleria mellonella (wax moth) larvae were evaluated as a surrogate host; we found that B. pseudomallei and B. mallei, but not other phylogenetically related bacteria, were highly pathogenic for this insect. More importantly, four previously characterized B. mallei mutants with reduced virulence in hamsters or mice had similarly reduced virulence in G. mellonella larvae. Site-specific inactivation of selected genes in the computationally derived virulome identified three new potential virulence genes, each of which was required for rapid and efficient killing of larvae. Thus, this approach may provide a means to quickly identify high-probability virulence genes in B. pseudomallei, B. mallei, and other pathogens.

Published ahead of print on 25 January 2008.

Supplemental material for this article may be found at http://jb.asm.org/.