Cloning and nucleotide sequence of the gene coding for the major 25-kilodalton outer membrane protein of Brucella abortus

Wergifosse, P de; Lintermans, P.; Limet, J N; Cloeckaert, Axel

doi:10.1128/jb.177.7.1911-1914.1995

Cited by 42 publications

(40 citation statements)

References 16 publications

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“…The genes omp25, omp31, and omp2b (which encode the major 25-, 31-, and 36-kDa Omps, respectively) and the genes pal, omp10, omp19, and omp1 (which encode the 16-, 10-, 19-, and 89-kDa minor Omps, respectively) have been cloned and sequenced (10,16,18,28,31,43,44,50; Bearden and Ficht, GenBank accession number U51683). Omp2b functions as a porin, and the 16-kDa Omp shows significant similarity to the peptidoglycan-associated lipoproteins Pal of gram-negative bacteria (32,44).…”

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Effect of omp10 or omp19 Deletion on Brucella abortus Outer Membrane Properties and Virulence in Mice

et al. 2002

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The distinctive properties of Brucella outer membrane have been considered to be critical for Brucella sp. virulence. Among the outer membrane molecules possibly related to these properties, Omp10 and Omp19 are immunoreactive outer membrane lipoproteins. Moreover, these proteins of Brucella could constitute a new family of outer membrane proteins specifically encountered in the family Rhizobiaceae. We evaluated the impact of omp10 or omp19 deletion on Brucella abortus outer membrane properties and virulence in mice. The omp10 mutant was dramatically attenuated for survival in mice and was defective for growth in minimal medium but was not impaired in intracellular growth in vitro, nor does it display clear modification of the outer membrane properties. Significantly fewer brucellae were recovered from the spleens of mice infected with the omp19 mutant than from those of mice infected with the parent strain at 4 and 8 weeks postinfection. The omp19 mutant exhibited an increase in sensitivity to the polycation polymyxin B and to sodium deoxycholate. These results indicate that inactivation of the omp19 gene alters the outer membrane properties of B. abortus.Brucellae are gram-negative bacteria that cause human disease and significant worldwide economic losses due to infection of livestock. These bacteria are able to multiply within professional and nonprofessional phagocytes, but the exact mechanisms whereby Brucella spp. intracellularly parasitize the host are still to be defined (4,5,7,15,30,36,37).The Brucella outer membrane has been proposed to be involved in virulence (i.e., resistance to bactericidal cationic peptides and polycations, permeability to hydrophobic agents, resistance to divalent cation chelators, and poor activation of bactericidal mechanisms by lipopolysaccharide [LPS]) (for a review, see reference 35). Correlation between these properties and specific surface molecules can be studied by genetically engineering mutations in the genes and determining the resultant phenotype.The analysis of genetically defined rough mutants of Brucella melitensis and Brucella abortus confirmed the involvement of lipopolysaccharide O side chain in Brucella in vivo survival (1,24,25,34,45,46). The recently identified BvrR-BvrS twocomponent regulatory system is involved in the control of outer membrane properties such as resistance to bactericidal polycations and is highly relevant for the virulence of B. abortus (40).The molecular characterization of several Brucella outer membrane proteins (Omps) has been reported over the past years. The genes omp25, omp31, and omp2b (which encode the major 25-, 31-, and 36-kDa Omps, respectively) and the genes pal, omp10, omp19, and omp1 (which encode the 16-, 10-, 19-, and 89-kDa minor Omps, respectively) have been cloned and sequenced (10,

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Effect of omp10 or omp19 Deletion on Brucella abortus Outer Membrane Properties and Virulence in Mice

et al. 2002

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“…it was able to protect mice against intranasal challenge [12], and 36 kDa [13] OMPs; and B. ovis the upper liquid was collected into a 1.5 ml tube and 25 kDa OMP [14]. The molecular sizes of these OMPs added 630 ýd isopropanol.…”

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Cloning, expression, and purification of Brucella suis outer membrane proteins

Ding

Bhattacharjee

Nikolich

et al. 2005

Protein Expression and Purification

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“…The gene coding for Omp25 of B. abortus (omp25) was recently cloned and sequenced [26]. Polymerase chain reaction and restriction fragment length polymorphism analysis showed that the omp25 gene is highly conserved among BrucelZa spp.…”

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Evaluation Of Immunogenicity and Protective Activity in Balb/C Mice of the 25-kDa Major Outer-Membrane Protein Of Brucella Melitensis (Omp25) Expressed in Escherichia Coli

Bowden

Cloeckaert²,

Zygmunt³

et al. 1998

Journal of Medical Microbiology

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The antibody response specific to the 25-kDa major outer-membrane protein (Omp25) of Brucella melitensis expressed in Escherichia coli was assessed in BALB/c mice. Groups of mice were immunised and boosted either with sonicated E. coli carrying plasmid pAC2533 -E. coli (pAC2533) -expressing the gene coding for Omp25 (omp25 gene) of B. melitensis, or with E. coli carrying plasmid pUCl9 -E. coli (pUC19). One control group received saline. The evolution of antibody responses was investigated by indirect ELISA with whole rough (R) B. melitensis H38 cells as antigen. Serum antibody titres of mice immunised with E. coli (pAC2533) were appreciably higher than those of mice immunised with E. coli (pUC19). The specificity to Omp25 of murine antibodies induced by E. coli (pAC2533) was demonstrated by SDS-PAGE and immunoblotting of five B. melitensis strains. Binding of antibody in E. coli (pAC2533) immune sera to the surface of B. melitensis strains differing in their smooth lipopolysaccharide (S-LPS) expression was also studied by whole-cell ELISA and by flow cytometry. Antibody reactivity to R and smooth-rough (S-R) was much stronger than that to smooth (S) B. melitensis strains, indicating a much better accessibility of Omp25 to antibody on strains lacking or expressing less O-polysaccharide on their surface. The antibodies to Omp25 were predominantly of IgG2a isotype. The capacity of E. coli (pAC2533) to induce protective immune responses against four challenge strains of B. melitensis was further evaluated in mice. Significant reductions in splenic infections, in comparison with mice immunised with E. coli (pUC19) and unimmunised (saline injection) mice, were observed in R B. melitensis B115, S-R B. melitensis EP and S B. melitensis H38 infected mice. Protection against S B. melitensis 16M was not significant. The data from the present study, together with previous results, suggest that humoral immunity against probably conformational, well-exposed epitopes of the Omp25 could contribute to protective mechanisms against B. melitensis infection in mice.

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Cloning and nucleotide sequence of the gene coding for the major 25-kilodalton outer membrane protein of Brucella abortus

Cited by 42 publications

References 16 publications

Effect of omp10 or omp19 Deletion on Brucella abortus Outer Membrane Properties and Virulence in Mice

Effect of omp10 or omp19 Deletion on Brucella abortus Outer Membrane Properties and Virulence in Mice

Cloning, expression, and purification of Brucella suis outer membrane proteins

Evaluation Of Immunogenicity and Protective Activity in Balb/C Mice of the 25-kDa Major Outer-Membrane Protein Of Brucella Melitensis (Omp25) Expressed in Escherichia Coli

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