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Infection and Immunity, November 1998, p. 5485-5493, Vol. 66, No. 11
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Identification of the Perosamine Synthetase Gene of Brucella melitensis 16M and Involvement of Lipopolysaccharide O Side Chain in Brucella Survival in Mice and in Macrophages

Fabrice Godfroid,1,* Bernard Taminiau,1 Isabelle Danese,1 Philippe Denoel,1 Anne Tibor,1 Vincent Weynants,1 Axel Cloeckaert,2 Jacques Godfroid,3 and Jean-Jacques Letesson1

Unité de Recherche en Biologie Moléculaire (URBM), Laboratoire d'Immunologie et de Microbiologie, Facultés Universitaires Notre Dame de la Paix, Namur,1 and Centre d'Étude et de Recherche Vétérinaire et Agrochimique (CERVA), Brussels,3 Belgium, and Institut National de la Recherche Agronomique, Centre de Recherche de Tours, Nouzilly, France2

Received 2 March 1998/Returned for modification 27 May 1998/Accepted 14 August 1998

Brucella organisms are facultative intracellular bacteria that may infect many species of animals as well as humans. The smooth lipopolysaccharide (S-LPS) has been reported to be an important virulence factor of these organisms, but the genetic basis of expression of the S-LPS O antigen has not yet been described. Likewise, the role of the O side chain of S-LPS in the survival of Brucella has not been clearly defined. A mini-Tn5 transposon mutant library of Brucella melitensis 16M was screened by enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies (MAbs) directed against the O side chain of Brucella. One mutant, designated B3B2, failed to express any O side chain as confirmed by ELISA, Western blot analysis, and colony coloration with crystal violet. Nucleotide sequence analysis demonstrated that the transposon disrupted an open reading frame with significant homology to the putative perosamine synthetase genes of Vibrio cholerae O1 and Escherichia coli O157:H7. The low G+C content of this DNA region suggests that this gene may have originated from a species other than a Brucella sp. The survival of B. melitensis mutant strain B3B2 in the mouse model and in bovine macrophages was examined. The results suggested that S-LPS or, more precisely, its O side chain is essential for survival in mice but not in macrophages.


* Corresponding author. Mailing address: Unité de Recherche en Biologie Moléculaire (URBM), Laboratoire d'Immunologie et de Microbiologie, Facultés Universitaire Notre Dame de la Paix, 61 rue de Bruxelles, B-5000, Namur, Belgium. Phone: 32-81-72 44 03. Fax: 32-81-72 44 20. E-mail: Fabrice.Godefroid{at}fundp.ac.be.


Infection and Immunity, November 1998, p. 5485-5493, Vol. 66, No. 11
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.



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