羊种布鲁氏菌M5-90 omp31蛋白原核表达  

Cloning and Prokaryotic Expression of Omp31 Gene from Brucella M5-90

作　　者：张红星[1] 陈创夫[2] 王远志[3] 唐利燕[2] 

机构地区：[1]石河子大学生命科学院,石河子832003 [2]石河子大学动物科技学院,石河子832003 [3]石河子大学医学院,石河子832002

出　　处：《石河子大学学报（自然科学版）》2009年第5期593-596,共4页Journal of Shihezi University(Natural Science)

基　　金：973计划前期研究专项(2007CB116309);国际科技合作项目(2006DFA33740);国家自然科学基金(30760187);国家自然科学基金项目(30800813)

摘　　要：克隆、测序布鲁氏菌疫苗株M5-90外膜蛋白基因OMP31,原核表达OMP31并对其检测。从羊种布鲁氏菌M5-90中PCR获得OMP31基因,连接到pBS-T克隆质粒并测序;将测序正确的基因片段克隆入大肠杆菌表达载体pET-28a,进行SDS-PAGE,而后Western-blot检测。结果M5-90疫苗株的OMP31基因序列与羊种布鲁氏菌参考株16M的同源性为99.03%;外膜蛋白基因OMP31在大肠杆菌表达后能够被Western-blot检测到。结论:表达的布鲁氏菌疫苗株M5-90外膜蛋白OMP31可以被布鲁氏菌特异性血清识别,表现出良好的抗原性,为以后实验室诊断和疫苗的研究做好坚实的上游工作。Brucella melitensis M5-90 vaccine strain OMP31 gene was cloned, sequenced,expressed and detected. OMP31 gene was amplified by PCR,cloned into pBS-T vector and sequenced. The objective gene was recombined into pET-28a vector, expressed and detected by SDS-PAGE and Western-blot. The results showed there was 99.03% identity for OMP31 gene between B. melitensis M5-90 vaccine strain and 16M reference strain and the expressed OMP31 was detected by Western-blot. The conclusion conveyed us that the special serum against bruceUa wild strain could recognize the expressed OMP31 and show it is an immunodominant antigen. This experiment gives us a good upstream work to laboratory diagnosis and study of vaccine.

关 键 词：羊种布鲁氏菌疫苗株M5-90 omp31基因 克隆 测序 Western—blot检测 

分 类 号：S852.614[农业科学—基础兽医学]