Murray, Gerald L., Morel, Viviane, Cerqueira, Gustavo M., Croda, Julio, Srikram, Amporn, Henry, Rebekah, Ko, Albert I., Dellagostin, Odir A., Bulach, Dieter M., Sermswan, Rasana W., Adler, Ben, and Picardeau, Mathieu (2009) Genome-wide transposon mutagenesis in pathogenic Leptospira species. Infection and Immunity, 77 (2). pp. 810-816.

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Abstract

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.

Item ID:	37427
Item Type:	Article (Research - C1)
ISSN:	1098-5522
Funders:	National Health and Medical Research Council of Australia (NHMRC), Australian Research Council (ARC), Institut Pasteur, French Ministry of Research (FMR), Fiocruz-Pasteur Scientific Cooperation Agreement, Brazilian National Research Foundation (CNPq), National Institute of Health (NIH), USA
Projects and Grants:	FMR ANR Jeunes Chercheurs no. 05-JCJC-0105-01, CNPq Instituto Milenio 420067/2005, NIH 5 R01 AI052473 , NIH 2 R01 AI034431, NIH 2 D43 TW-00919
Date Deposited:	19 Mar 2015 01:40
FoR Codes:	06 BIOLOGICAL SCIENCES > 0605 Microbiology > 060502 Infectious Agents @ 100%
SEO Codes:	97 EXPANDING KNOWLEDGE > 970106 Expanding Knowledge in the Biological Sciences @ 100%
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