Identification and characterization of in vivo attenuated mutants of Brucella melitensis.

Animals; Bacterial Proteins/genetics/metabolism; Brucella melitensis/genetics/growth & development/pathogenicity; Brucellosis/microbiology; DNA Transposable Elements/genetics; Gene Library; Hela Cells; Humans; Macrophages/microbiology; Mice; Mice, Inbred BALB C; Mutagenesis, Insertional; Virulence/genetics

Abstract :

[en] Brucella melitensis 16M is a Gram-negative alpha2-proteobacterium responsible for abortion in goats and for Malta fever in humans. This facultative intracellular pathogen invades into and survives within both professional and non-professional phagocytes. Signature-tagged mutagenesis (STM) was used to identify genes required for the in vivo pathogenesis of Brucella. A library of transposon mutants was screened in a murine infection model. Out of 672 mutants screened, 20 were not recovered after a 5 day passage in BALB/c mice. The attenuation of 18 mutants was confirmed using an in vivo competition assay against the wild-type strain. The 18 mutants were characterized further for their ability to replicate in murine macrophages and in HeLa cells. The sequences disrupted by the transposon in the mutants have homology to genes coding for proteins of different functional classes: transport, amino acid and DNA metabolism, transcriptional regulation, peptidoglycan synthesis, a chaperone-like protein and proteins of unknown function. The mutants selected in this study provide new insights into the molecular basis of Brucella virulence.

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Lestrate, P.

Delrue, R. M.

DANESE, Isabelle ; Université de Liège - ULiège > GIGA-M : Coordination scientifique

Didembourg, C.

Taminiau, Bernard ; Université de Liège - ULiège > Département de sciences des denrées alimentaires > Microbiologie des denrées alimentaires

Fonck, P.

De Bolle, X.

Tibor, A.

Tang, C. M.

Letesson, J. J.

Language :

English

Title :

Identification and characterization of in vivo attenuated mutants of Brucella melitensis.

Publication date :

2000

Journal title :

Molecular Microbiology

ISSN :

0950-382X

eISSN :

1365-2958

Publisher :

Blackwell, Oxford, United Kingdom

Volume :

Issue :

Pages :

543-51

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 10 January 2013

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135

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118

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129

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