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Genome-Wide Transposon Mutagenesis in Pathogenic Leptospira Species▿ ‡

Identifieur interne : 000491 ( Ncbi/Checkpoint ); précédent : 000490; suivant : 000492

Genome-Wide Transposon Mutagenesis in Pathogenic Leptospira Species▿ ‡

Auteurs : Gerald L. Murray ; Viviane Morel ; Gustavo M. Cerqueira ; Julio Croda ; Amporn Srikram ; Rebekah Henry ; Albert I. Ko ; Odir A. Dellagostin ; Dieter M. Bulach ; Rasana W. Sermswan ; Ben Adler ; Mathieu Picardeau

Source :

Infection and Immunity [ 0019-9567 ] ; 2008.

RBID : PMC:2632054

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.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2632054

DOI: 10.1128/IAI.01293-08
PubMed: 19047402
PubMed Central: 2632054

Affiliations:

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PMC:2632054

Le document en format XML

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 Species<xref ref-type="fn" rid="fn3">▿</xref>
 <xref ref-type="fn" rid="fn2">‡</xref>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Genome-Wide Transposon Mutagenesis in Pathogenic <italic>Leptospira</italic>
 Species<xref ref-type="fn" rid="fn3">▿</xref>
 <xref ref-type="fn" rid="fn2">‡</xref>
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<front><div type="abstract" xml:lang="en"><p><italic>Leptospira interrogans</italic>
 is the most common cause of leptospirosis in humans and animals. Genetic analysis of <italic>L. interrogans</italic>
 has been severely hindered by a lack of tools for genetic manipulation. Recently we developed the <italic>mariner</italic>
-based transposon <italic>Himar1</italic>
 to generate the first defined mutants in <italic>L. interrogans</italic>
. 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 <italic>L. interrogans</italic>
 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 <italic>L. interrogans</italic>
. This library provides a valuable resource for the study of gene function in <italic>L. interrogans</italic>
. Combined with the genome sequences of <italic>L. interrogans</italic>
, this provides an opportunity to investigate genes that contribute to pathogenesis and will provide a better understanding of the biology of <italic>L. interrogans</italic>
.</p>
</div>
</front>
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<name sortKey="Cerqueira, Gustavo M" sort="Cerqueira, Gustavo M" uniqKey="Cerqueira G" first="Gustavo M." last="Cerqueira">Gustavo M. Cerqueira</name>
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<name sortKey="Dellagostin, Odir A" sort="Dellagostin, Odir A" uniqKey="Dellagostin O" first="Odir A." last="Dellagostin">Odir A. Dellagostin</name>
<name sortKey="Henry, Rebekah" sort="Henry, Rebekah" uniqKey="Henry R" first="Rebekah" last="Henry">Rebekah Henry</name>
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<name sortKey="Morel, Viviane" sort="Morel, Viviane" uniqKey="Morel V" first="Viviane" last="Morel">Viviane Morel</name>
<name sortKey="Murray, Gerald L" sort="Murray, Gerald L" uniqKey="Murray G" first="Gerald L." last="Murray">Gerald L. Murray</name>
<name sortKey="Picardeau, Mathieu" sort="Picardeau, Mathieu" uniqKey="Picardeau M" first="Mathieu" last="Picardeau">Mathieu Picardeau</name>
<name sortKey="Sermswan, Rasana W" sort="Sermswan, Rasana W" uniqKey="Sermswan R" first="Rasana W." last="Sermswan">Rasana W. Sermswan</name>
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Genome-Wide Transposon Mutagenesis in Pathogenic Leptospira Species▿ ‡

Genome-Wide Transposon Mutagenesis in Pathogenic Leptospira Species▿ ‡

Source :

Abstract

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Links to Exploration step

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

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Pour générer des pages wiki