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Infection and Immunity, February 2009, p. 810-816, Vol. 77, No. 2
0019-9567/09/$08.00+0     doi:10.1128/IAI.01293-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Genome-Wide Transposon Mutagenesis in Pathogenic Leptospira Species{triangledown} ,{ddagger}

Gerald L. Murray,1 Viviane Morel,2 Gustavo M. Cerqueira,2,3 Julio Croda,2,4 Amporn Srikram,5 Rebekah Henry,1 Albert I. Ko,4,6 Odir A. Dellagostin,3 Dieter M. Bulach,1,{dagger} Rasana W. Sermswan,7 Ben Adler,1,8* and Mathieu Picardeau2*

Australian Bacterial Pathogenesis Program, Department of Microbiology, Monash University, Clayton, VIC 3800, Australia,1 Institut Pasteur, Unité de Biologie des Spirochètes, Paris, France,2 Centro de Biotecnologia, Universidade Federal de Pelotas, P.O. Box 354, 96010-900, Pelotas, RS, Brazil,3 Gonçalo Moniz Research Center, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Salvador, Brazil,4 Melioidosis Research Center, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand,5 Division of International Medicine and Infectious Disease, Weill Medical College of Cornell University, New York, New York,6 Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand,7 Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, Department of Microbiology, Monash University, Clayton, VIC 3800, Australia8

Received 21 October 2008/ Returned for modification 11 November 2008/ Accepted 23 November 2008

Leptospira interrogans is the most common cause of leptospirosis in humans and animals. Genetic analysis of L. interrogans has been severely hindered by a lack of tools for genetic manipulation. Recently we developed the mariner-based transposon Himar1 to generate the first defined mutants in L. interrogans. In this study, a total of 929 independent transposon mutants were obtained and the location of insertion determined. Of these mutants, 721 were located in the protein coding regions of 551 different genes. While sequence analysis of transposon insertion sites indicated that transposition occurred in an essentially random fashion in the genome, 25 unique transposon mutants were found to exhibit insertions into genes encoding 16S or 23S rRNAs, suggesting these genes are insertional hot spots in the L. interrogans genome. In contrast, loci containing notionally essential genes involved in lipopolysaccharide and heme biosynthesis showed few transposon insertions. The effect of gene disruption on the virulence of a selected set of defined mutants was investigated using the hamster model of leptospirosis. Two attenuated mutants with disruptions in hypothetical genes were identified, thus validating the use of transposon mutagenesis for the identification of novel virulence factors in L. interrogans. This library provides a valuable resource for the study of gene function in L. interrogans. Combined with the genome sequences of L. interrogans, this provides an opportunity to investigate genes that contribute to pathogenesis and will provide a better understanding of the biology of L. interrogans.

* Mailing address for Mathieu Picardeau: Unité de Biologie des Spirochètes, Institut Pasteur, 28 rue du docteur Roux, 75724 Paris Cedex 15, France. Phone: 33 (1) 45 68 83 68. Fax: 33 (1) 40 61 30 01. E-mail: mpicard{at}pasteur.fr. Mailing address for Ben Adler: Department of Microbiology, Monash University, Wellington Road, Victoria 3800, Australia. Phone: 61 3 9905 4815. Fax: 61 3 9905 4811. E-mail: ben.adler{at}med.monash.edu.au

{triangledown} Published ahead of print on 1 December 2008.

{ddagger} Supplemental material for this article may be found at http://iai.asm.org/.

Editor: A. J. Bäumler

{dagger} Present address: CSIRO Livestock Industries, Australian Animal Health Laboratory (AAHL), Geelong, VIC 3220, Australia.

Infection and Immunity, February 2009, p. 810-816, Vol. 77, No. 2
0019-9567/09/$08.00+0     doi:10.1128/IAI.01293-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Murray, G. L., Srikram, A., Hoke, D. E., Wunder, E. A. Jr., Henry, R., Lo, M., Zhang, K., Sermswan, R. W., Ko, A. I., Adler, B. (2009). Major Surface Protein LipL32 Is Not Required for Either Acute or Chronic Infection with Leptospira interrogans. Infect. Immun. 77: 952-958 [Abstract] [Full Text]  


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