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In the present report, we have extended this study to the second etiologic agent in ovine brucellosis, <jats:italic>B. ovis<\/jats:italic>, responsible for ram epididymitis and infertility. The combination of 2\u2010D gel electrophoresis and protein microsequencing facilitated the location and identification of the major proteins of <jats:italic>B. ovis<\/jats:italic> on the 2\u2010D pattern. These proteins comprised cytoplasmic, periplasmic and some membrane proteins except the major outer membrane proteins. By comparing 2\u2010D gel profiles of <jats:italic>B. ovis<\/jats:italic> with that of <jats:italic>B. melitensis<\/jats:italic> described previously, a few proteins with different expression levels were readily identified. Serum from a ram naturally infected with <jats:italic>B. ovis<\/jats:italic> was used in immunoblotting studies to identify immunogenic proteins recognized during the course of infection. This serum showed antibody reactivity against approximately 82 protein spots. Twenty\u2010one of these proteins were identified either by use of monoclonal antibodies or by <jats:italic>N<\/jats:italic>\u2010terminal microsequencing. Several proteins previously described in earlier <jats:italic>Brucella<\/jats:italic> works were identified: the 89 kDa outer membrane protein, DnaK, GroEL, BP26, and Cu\u2010Zn superoxide dismutase. Eight proteins had amino acid sequences homologous to those of various proteins from other bacteria found in protein databases: NikA, dihydrolipoamide succinyltransferase, a hypothetical 31 kDa protein, malate dehydrogenase, succinyl\u2010CoA synthetase alpha subunit, an amino acid ABC type transporter, Leu\/ Ile\/ Val\u2010binding protein precursor, and ClpP. The remaining eight proteins had <jats:italic>N<\/jats:italic>\u2010terminal sequences lacking similarity to existing database entries. Thus, the 2\u2010D PAGE analysis provided a convenient first approach in the characterization of immunogenic proteins.<\/jats:p>","DOI":"10.1002\/elps.1150180824","type":"journal-article","created":{"date-parts":[[2005,5,28]],"date-time":"2005-05-28T12:14:38Z","timestamp":1117282478000},"page":"1491-1497","source":"Crossref","is-referenced-by-count":49,"title":["Identification and characterization of <i>Brucella ovis<\/i> immunogenic proteins using two\u2010dimensional electrophoresis and immunoblotting"],"prefix":"10.1002","volume":"18","author":[{"given":"Ana P.","family":"Teixeira\u2010Gomes","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Axel","family":"Cloeckaert","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guy","family":"B\u00e9zard","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ra\u00fal A.","family":"Bowden","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G\u00e9rard","family":"Dubray","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michel S.","family":"Zygmunt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2005,4,14]]},"reference":[{"key":"e_1_2_1_2_2","first-page":"377","volume-title":"Bergey's manual of systematic bacteriology","author":"Corbel M. 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