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
Department of Bacterial Diseases, Walter Reed Army Institute of Research, Washington, DC 20307-5100, USA.
Infect Immun. 1996 Jul;64(7):2490-9. doi: 10.1128/iai.64.7.2490-2499.1996.
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 lambdagt11 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.
布鲁氏菌3组抗原（Ags）是分子量在25至30 kDa之间的外膜蛋白（OMPs）。这些OMPs之所以受到关注，部分原因在于它们有潜力用作疫苗和诊断试剂。我们利用人类恢复期抗体（Ab）从羊种布鲁氏菌16M基因组DNA的λgt11文库中克隆了一个编码28-kDa蛋白的基因。DNA序列分析揭示了一个单一的开放阅读框，其编码的蛋白分子量为26,552 Da。该28-kDa蛋白的一级氨基酸序列与先前描述的流产布鲁氏菌3组Ag、Omp25有43%的相似性（P. de Wergifosse、P. Lintermans、J. N. Limet和A. Cloeckaert，《细菌学杂志》177:1911 - 1914，1995年）。与已知的3组OMP的相似性、与针对流产布鲁氏菌Ags制备的Ab的免疫反应性、全细胞的免疫标记以及Southern杂交，使我们得出结论：羊种布鲁氏菌的28-kDa蛋白是一种与流产布鲁氏菌Omp25不同的3组蛋白。我们将羊种布鲁氏菌的该蛋白命名为Omp28。感染流产布鲁氏菌和猪布鲁氏菌的患者的人类恢复期血清以及针对灭活的流产布鲁氏菌全细胞制备的兔抗血清，在Western印迹（免疫印迹）中能识别羊种布鲁氏菌Omp28。此外，感染羊种布鲁氏菌光滑菌株的小鼠和山羊产生了针对Omp28的Ab。我们的结果或许能开始解释在布鲁氏菌Ags中观察到的分子量变异性，并指出它们在预防感染疫苗接种以及疾病诊断中的可能用途。
