UNESCO-MIRCEN for Marine Biotechnology, Department of Fishery Microbiology, Karnataka Veterinary, Animal & Fisheries Sciences University, College of Fisheries, Mangalore, India.
Microbiol Res. 2011 Dec 20;167(1):1-7. doi: 10.1016/j.micres.2011.02.002. Epub 2011 Apr 8.
Outer membrane protein A (OmpA) is a component of the outer membrane of Edwardsiella tarda and is wildly distributed in Enterobacteriaceae family. The gene encoding the OmpA protein was cloned from E. tarda and expressed in Escherichia coli M15 cells. The recombinant OmpA protein containing His(6) residues was estimated to have a molecular weight of ~38kDa. In Western blot the native protein showed expression at ~36kDa molecular weight which was within the range of major outer membrane proteins (36-44kDa) observed in this study. All E. tarda isolates tested harbored the ompA gene and the antibody raised to this protein was seen to cross react with other Gram negative bacteria. The OmpA protein characterized in this study was observed to be highly immunogenic in both rabbit and fish. In Enzyme linked immunosorbent assay, rabbit antisera showed an antibody titer of 1: 128,000. Common carp vaccinated with recombinant OmpA protein elicited high antibody production and immunized fish showed a relative percentage survival of 54.3 on challenge.
外膜蛋白 A(OmpA)是迟缓爱德华氏菌外膜的组成部分,广泛分布于肠杆菌科。从迟缓爱德华氏菌中克隆了编码 OmpA 蛋白的基因,并在大肠杆菌 M15 细胞中表达。含有 His(6)残基的重组 OmpA 蛋白估计分子量约为 38kDa。在 Western blot 中,天然蛋白在 ~36kDa 分子量处表达,这在本研究中观察到的主要外膜蛋白(36-44kDa)范围内。所有测试的迟缓爱德华氏菌分离株都携带 ompA 基因,针对该蛋白产生的抗体与其他革兰氏阴性菌发生交叉反应。本研究中鉴定的 OmpA 蛋白在兔和鱼中均具有高度免疫原性。在酶联免疫吸附试验中,兔抗血清的抗体效价为 1:128,000。用重组 OmpA 蛋白免疫的鲤鱼产生了高抗体产生,免疫鱼在攻毒后相对存活率为 54.3%。