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Infection and Immunity, April 2001, p. 2054-2065, Vol. 69, No. 4
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.
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
*
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.
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