内蒙古分离株M3253羊种布鲁菌omp31基因的表达及抗原性分析

Expression and antigenicity analysis of the omp31 gene from Brucella melitensis strain M3253 isolated from Inner mongolia

为了在大肠杆菌中表达羊种布鲁菌omp31基因并鉴定重组蛋白的抗原性,试验采用PCR技术从内蒙古分离株M3253羊种布鲁菌中扩增得到布鲁菌omp31基因片段,插入pMD18-T载体克隆测序,并将目的基因插入原核表达载体pET-30a中,构建重组质粒pET-30a-omp31,在大肠杆菌BL21(DE3)中表达该基因,并用SDS-PAGE和Western-blot检测该蛋白。结果表明:成功构建了重组质粒pET-30a-omp31,并在大肠杆菌中成功表达了omp31基因,融合蛋白与布鲁菌阳性血清发生特异性反应。说明布鲁菌omp31基因成功表达,重组蛋白可与羊种布鲁菌阳性血清发生特异性反应。

To express the omp31 gene of Brucella in E.coli and identify the antigenicity of the recombinant protein omp31 of Brucella.The omp31 gene were got from Brucella melitensis isolated in inner mongolia by polymerase chain reaction（PCR） and cloned into pMD18-T.After sequencing,the omp31 gene were cloned into vector pET-30a and expressed in E.coli BL21（DE3）,then detected by SDS-PAGE and Western-blot.The results showed that the recombinant plasmid pET-30a-omp31 was constructed and expressed successfully in E.coli.It indicates that the recombinant protein can react with Brucella melitensis positive serum,it provides a good foundation for the study of laboratory diagnosis and preparation of the vaccine.