Article (Scientific journals)
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.
Godfroid, F.; Taminiau, Bernard; DANESE, Isabelle et al.
1998In Infection and Immunity, 66 (11), p. 5485-93
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Keywords :
Amino Acid Sequence; Animals; Base Sequence; Brucella melitensis/genetics/growth & development; Brucellosis/enzymology/genetics/microbiology; Carbohydrate Epimerases/genetics; Cattle; Cell Line, Transformed; Cloning, Molecular; Disease Models, Animal; Genes, Bacterial/immunology; Lipopolysaccharides/chemistry/immunology; Macrophages, Peritoneal/enzymology/microbiology; Mice; Molecular Sequence Data; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Transaminases/genetics
Abstract :
[en] 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.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Godfroid, F.
Taminiau, Bernard  ;  Université de Liège - ULiège > Département de sciences des denrées alimentaires > Microbiologie des denrées alimentaires
DANESE, Isabelle ;  Université de Liège - ULiège > GIGA-M : Coordination scientifique
Denoel, P.
Tibor, A.
Weynants, V.
Cloeckaert, A.
Godfroid, J.
Letesson, J. J.
Language :
English
Title :
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.
Publication date :
1998
Journal title :
Infection and Immunity
ISSN :
0019-9567
eISSN :
1098-5522
Publisher :
American Society for Microbiology, Washington, United States - District of Columbia
Volume :
66
Issue :
11
Pages :
5485-93
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 10 January 2013

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