布鲁氏菌Cu/Zn-SOD基因的原核表达及其免疫原性分析  

Prokaryotic expression of Brucella melitensis Cu/Zn-SOD gene and analysis of its immunogenicity

作　　者：覃晓琳[1] 乔祖健[2] 胡森[2] 刘文兴[2] 赵云[2] 李继昌[1] 步志高[2] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/农业部兽医公共卫生重点开放实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2013年第4期317-320,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：农业公益性行业科研专项(200803018)

摘　　要：为分析布鲁氏菌超氧化物歧化酶(Cu/Zn-SOD)蛋白的免疫原性,本研究将布鲁氏菌强毒株M28的Cu/Zn-SOD基因经PCR扩增并克隆至pET-32a(+)中进行重组表达和纯化。经western blot分析表明,重组蛋白SOD(rSOD)可以与布鲁氏菌自然感染血清发生特异性反应。同时,以rSOD免疫BALB/c小鼠制备的多抗血清可以与布鲁氏菌疫苗株M5-90发生特异性反应;以rSOD为包被抗原进行间接ELISA检测,结果显示rSOD能够区分布鲁氏菌阳性血清和阴性血清。因此,rSOD蛋白具有良好的免疫原性和ELISA反应原性,为利用rSOD蛋白建立布鲁氏菌检测方法奠定基础。To evaluate immunogenicity of Brucella melitensis Cu-Zn superoxide dismutase （Cu/Zn-SOD）, the gene coding Cu/Zn-SOD was amplified by PCR and subcloned into pET-32a for expression in E.coli. Western blot analysis showed that the expressed recombinant protein （rSOD） was about 35 ku, mainly in the form of inclusion body, which reacted with positive sera of B.melitensis detected by SDS-PAGE and western blot. Furthermore, the antiserum was prepared in mice by immunization of the purified recombinant protein. Western blot analysis demonstrated that the antiserum showed specific reaction with B.melitensis M5-90. An indirect ELISA was set up using purified rSOD as coating antigen. A total of 21 serum samples were examined, which showed that 13 serum samples were B.melitensis positive. In conclusion, the rSOD protein was good immunogenicity and could be applied for the detection of B.melitensis infection.

关 键 词：布鲁氏菌 Cu ZN SOD基因 原核表达 免疫原性 

分 类 号：S852.61[农业科学—基础兽医学]