This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Cuccui, J.
Right arrow Articles by Wren, B. W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Cuccui, J.
Right arrow Articles by Wren, B. W.

 Previous Article  |  Next Article 

Infection and Immunity, March 2007, p. 1186-1195, Vol. 75, No. 3
0019-9567/07/$08.00+0     doi:10.1128/IAI.01240-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Development of Signature-Tagged Mutagenesis in Burkholderia pseudomallei To Identify Genes Important in Survival and Pathogenesis{triangledown}

J. Cuccui,1 A. Easton,1 K. K. Chu,1 G. J. Bancroft,1 P. C. F. Oyston,2 R. W. Titball,2 and B. W. Wren1*

Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, Keppel Street, London WC1E 7HT,1 Defence Science and Technology Laboratory, Porton Down, Salisbury, Wiltshire SP4 0JQ, United Kingdom2

Received 4 August 2006/ Returned for modification 23 September 2006/ Accepted 4 December 2006

Burkholderia pseudomallei, the causative agent of melioidosis, is an important human pathogen in Southeast Asia and northern Australia for which a vaccine is unavailable. A panel of 892 double signature-tagged mutants was screened for virulence using an intranasal BALB/c mouse model of infection. A novel DNA tag microarray identified 33 mutants as being attenuated in spleens, while 6 were attenuated in both lungs and spleens. The transposon insertion sites in spleen-attenuated mutants revealed genes involved in several stages of capsular polysaccharide biosynthesis and DNA replication and repair, a putative oxidoreductase, ABC transporters, and a lipoprotein that may be important in intercellular spreading. The six mutants identified as missing in both lungs and spleens were found to have insertions in recA involved in the SOS response and DNA repair; putative auxotrophs of leucine, threonine, p-aminobenzoic acid, and a mutant with an insertion in aroB causing auxotrophy for aromatic compounds were also found. Murine challenge studies revealed partial protection in BALB/c mice vaccinated with the aroB mutant. The refined signature-tagged mutagenesis approach developed in this study was used to efficiently identify attenuating mutants from this highly pathogenic species and could be applied to other organisms.

* Corresponding author. Mailing address: Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, University of London, Keppel Street, London WC1E 7HT, United Kingdom. Phone: 44 (0) 207 927 2288. Fax: 44 (0) 207 637 4314. E-mail: Brendan.Wren{at}lshtm.ac.uk.

{triangledown} Published ahead of print on 22 December 2006.

Editor: F. C. Fang

Infection and Immunity, March 2007, p. 1186-1195, Vol. 75, No. 3
0019-9567/07/$08.00+0     doi:10.1128/IAI.01240-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Gawronski, J. D., Wong, S. M. S., Giannoukos, G., Ward, D. V., Akerley, B. J. (2009). Tracking insertion mutants within libraries by deep sequencing and a genome-wide screen for Haemophilus genes required in the lung. Proc. Natl. Acad. Sci. USA 106: 16422-16427 [Abstract] [Full Text]  
  • Malinverni, J. C., Silhavy, T. J. (2009). An ABC transport system that maintains lipid asymmetry in the Gram-negative outer membrane. Proc. Natl. Acad. Sci. USA 106: 8009-8014 [Abstract] [Full Text]  
  • Harland, D. N., Chu, K., Haque, A., Nelson, M., Walker, N. J., Sarkar-Tyson, M., Atkins, T. P., Moore, B., Brown, K. A., Bancroft, G., Titball, R. W., Atkins, H. S. (2007). Identification of a LolC Homologue in Burkholderia pseudomallei, a Novel Protective Antigen for Melioidosis. Infect. Immun. 75: 4173-4180 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»