中文摘要：

研究羊布鲁菌外膜蛋白Omp25d基因的克隆,原核表达,以及纯化,根据羊布鲁菌M5株外膜蛋白Omp25d蛋白基因序列设计引物,扩增出大小约为650bp的目的基因片断,克隆入融合表达载体pGEX-4T-1,构建重组质粒pGEX-4T-1-Omp25d。在大肠杆菌中将该蛋白表达并用亲和层析法纯化。用Western-blot分析方法鉴定GST-Omp25d蛋白。结果成功地构建了pGEX-4T-1-Omp25d原核表达载体并在大肠杆菌中表达了Omp25d基因,纯化后所获得的融合蛋白与兔抗布鲁菌血清发生特异性反应。表明研究成功构建了pGEX-4T-1-Omp25d元和表达载体,并且在大肠杆菌中进行表达,纯化的融合蛋白具有良好的免疫原性。

英文摘要：

To clone and express the brucella outer membrane protein Omp25d,the gene of the protein was amplified from Brucella melitunsis with PCR method,then the PCR product was subcloned into vector pGEX-4T-1 and exprussed in E.coli.It is purified via affinity chromatography purification system.The purified protein is detected by Western-blot.According to the SDS-PAGE and Western-blot analysis,the recombinant plasmid pGEX-4T-1-Omp25d could be expressed successfully in E.coli and the fusion protein could react with serum from Brucella infected rabbit.The fusion gene encoding Omp25d protein is constructed and purified successfully and the good immunogenicity of the protein with high efficiency of expression was demonstrated.