马耳他布鲁菌omp25的原核表达与免疫原性检测

Expression and antigenicity analysis of the omp25 of Brucell melitensis strain

为在大肠杆菌中表达马耳他布鲁菌omp25基因并鉴定重组蛋白的抗原性,从马耳他布鲁菌中用聚合酶链反应技术(PCR)扩增得到布鲁菌omp25基因片段,并将目的基因插入原核表达载体pET-32a中,构建重组质粒pET-32a-omp25转入大肠杆菌Rosetta中表达,用SDS-PAGE和Western-blot检测表达蛋白。结果显示成功构建了重组质粒pET-32a-omp25,并在大肠杆菌Rosetta中获得了重组蛋白,重组蛋白与布鲁菌阳性血清发生特异性反应。表明重组质粒pET-32a-omp25可以在大肠杆菌Rosetta中成功表达,并且重组蛋白可与布鲁菌阳性血清发生特异性反应,说明该重组蛋白有良好的免疫原性,该研究为以后疫苗的研制及布鲁菌病的检测打下良好的基础。

To express omp25 gene of Brucella in E.coli Rosetta and determine the antigenicity of the recombinant protein omp25 of Brucella.The omp25 gene were obtained from Brucella melitensis by Polymerase chain reaction（PCR）,and the omp31 gene were cloned into vector pET-32a and expressed in E.coli Rosetta,then detected by SDS-PAGE and Western-blot.The recombinant plasmid pET-32a-omp31 was constructed successfully and the recombinant protein were expressed successfully in E.coli.The recombinant protein could react with Brucella melitensis positive serum.The recombinant protein can react with Brucella melitensis serum,which provides good foundation for the study of lab diagnosis and preparation of subunt vaccine.