Cloning of a Brucella melitensis group 3 antigen gene encoding Omp28, a protein recognized by the humoral immune response during human brucellosis. [Erratum: Oct 1996, v. 64 (10) p. 4414.]

Brucella group 3 antigens (Ags) are outer membrane proteins (OMPs) with a molecular mass ranging from 25 to 30 kDa. The OMPs are of interest partially because of their potential use as vaccine and diagnostic reagents. We used human convalescent antibody (Ab) to clone a gene that encoded a 28-kDa pro...

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Bibliographic Details
Published in:Infection and Immunity 1996-07, Vol.64 (7), p.2490-2499
Main Authors: Lindler, L.E, Hadfield, T.L, Tall, B.D, Snellings, N.J, Rubin, F.A, Van de verg, L.L, Hoover, D, Warren, R.L
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Antibodies, Bacterial - biosynthesis
Antibodies, Bacterial - blood
Antigens, Bacterial - genetics
Bacterial Outer Membrane Proteins - chemistry
Bacterial Outer Membrane Proteins - genetics
Bacterial Outer Membrane Proteins - immunology
Bacterial Vaccines - pharmacology
Bacteriology
Base Sequence
Biological and medical sciences
BRUCELLA MELITENSIS
Brucella melitensis - genetics
Brucella melitensis - immunology
BRUCELLOSE
Brucellosis - immunology
Brucellosis - prevention & control
BRUCELOSIS
Cloning, Molecular
DNA Primers - genetics
DNA, Bacterial - genetics
Fundamental and applied biological sciences. Psychology
Genes, Bacterial
Genetics
Goats
Humans
Mice
Microbiology
Molecular Sequence Data
Molecular Weight
Rabbits
REPONSE IMMUNITAIRE
RESPUESTA INMUNOLOGICA
Sequence Homology, Amino Acid
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Summary:Brucella group 3 antigens (Ags) are outer membrane proteins (OMPs) with a molecular mass ranging from 25 to 30 kDa. The OMPs are of interest partially because of their potential use as vaccine and diagnostic reagents. We used human convalescent antibody (Ab) to clone a gene that encoded a 28-kDa protein from a lambda gt11 library of Brucella melitensis 16M genomic DNA. DNA sequence analysis revealed a single open reading frame that would encode a protein of 26,552 Da. The 28-kDa protein had a primary amino acid sequence that was 43% similar to a previously described Brucella abortus group 3 Ag, Omp25 (P. de Wergifosse, P. Lintermans, J. N. Limet, and A. Cloeckaert, J. Bacteriol. 177:1911-1914, 1995). The similarity to a known group 3 OMP, immunoreactivity with Ab prepared against B. abortus group Ags, immunolabeling of whole cells, and Southern hybridization led to our conclusion that the B. melitensis 28-kDa protein was a group 3 protein distinct from B. abortus Omp25. We designated the B. melitensis protein Omp28. Human convalescent sera from patients infected with B. abortus and Brucella suis as well as rabbit antisera prepared against killed B. abortus whole cells recognized B. melitensis Omp28 on Western blots (immunoblots). Furthermore, mice and goats infected with smooth strains of B. melitensis produced Abs against Omp28. Our results may begin to explain the variability in molecular weight seen in Brucella group Ags and point toward their possible use in vaccination against infection as well as diagnosis of the disease
ISSN:0019-9567
1098-5522
DOI:10.1128/iai.64.7.2490-2499.1996