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Infection and Immunity, April 2001, p. 2054-2065, Vol. 69, No. 4
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.4.2054-2065.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Identification of New Genes Involved in the Virulence of Listeria monocytogenes by Signature-Tagged Transposon Mutagenesis

Nicolas Autret, Iharilalao Dubail, Patrick Trieu-Cuot, Patrick Berche, and Alain Charbit*

INSERM U-411, CHU Necker-Enfants Malades, 75730 Paris Cedex 15, France

Received 16 October 2000/Returned for modification 28 November 2000/Accepted 2 January 2001

Listeria monocytogenes is a gram-positive, facultative intracellular pathogen that can cause severe food-born infections in humans and animals. We have adapted signature-tagged transposon mutagenesis to L. monocytogenes to identify new genes involved in virulence in the murine model of infection. We used transposon Tn1545 carried on the integrative vector pAT113. Forty-eight tagged transposons were constructed and used to generate banks of L. monocytogenes mutants. Pools of 48 mutants were assembled, taking one mutant from each bank, injected into mice, and screened for those affected in their multiplication in the brains of infected animals. From 2,000 mutants tested, 18 were attenuated in vivo. The insertions harbored by these mutants led to the identification of 10 distinct loci, 7 of which corresponded to previously unknown genes. The properties of four loci involving putative cell wall components were further studied in vitro and in vivo. The data suggested that these components are involved in bacterial invasion and multiplication in the brain.

* Corresponding author. Mailing address: Faculté de Médecine Necker, 156, rue de Vaugirard, 75730 Paris Cedex 15, France. Phone: 33-1-40-61-53-76. Fax: 33-1-40-61-55-92. E-mail: charbit{at}necker.fr.

Infection and Immunity, April 2001, p. 2054-2065, Vol. 69, No. 4
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.4.2054-2065.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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