Cu/Zn SOD蛋白在布鲁菌S19疫苗株中的过表达及鉴定  

Overexpression and identification of Cu/Zn SOD in Brucella abortus strain 19

作　　者：邓雨明 段开放 侯欢欢 张燕[2] 彭其胜[1] DENG Yu-ming;DUAN Kai-fang;HOU Huan-huan;ZHANG Yan;PENG Qi-sheng(Key Laboratory for Zoonosis Re sea rch , Ministry of Education , hist it ate of Zoonosis,Jilin University , Changchun 130062 , China;Tumor Hospital of J ilin Province,Changchun 130021,China)

机构地区：[1]吉林大学人兽共患病研究所/教育部人兽共患病研究重点实验室,吉林长春130062 [2]吉林省肿瘤医院,吉林长春130021

出　　处：《中国兽医学报》2019年第4期711-715,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31372409)

摘　　要：以Cu/Zn SOD为目的基因,通过基因工程技术分别构建了组成型过表达系统pBBR-trc-sod和诱导型过表达系统pBBR-lacPtrc-sod,在布鲁菌S19疫苗株中进行了Cu/Zn SOD的过表达。同时,以大肠杆菌为宿主表达纯化的重组Cu/Zn SOD蛋白免疫家兔制备多克隆抗体,对2种过表达系统中Cu/Zn SOD蛋白的表达量进行分析。结果显示,诱导型Cu/Zn SOD过表达系统的表达量更高,且能与所制备的多抗发生特异性反应。结果表明,构建的过表达系统能够实现Cu/Zn SOD蛋白在布鲁菌S19疫苗株中过表达;同时,该试验也提示我们这种表达系统可应用于布鲁菌S19疫苗的抗原改造。Brucella abortus strain 19,as a live attenuated vaccine naturally attenuated from a virulent culture of B.abortus,has not been reported about its antigen modification at present.Due to the crucial role of Cu-Zn superoxide dismutase(Cu/Zn SOD) in immunoprotection of B.abortus strain 19,a constitutive Cu/Zn SOD expression system pBBR-trc-sod and an inducible Cu/Zn SOD expression system pBBR-lacPtrc-sod were constructed by genetic engineering in this research.Furthermore,these systems were used in overexpressing Cu/Zn SOD in B.abortus S19 for the first time.Meanwhile,the antiserum was prepared in mice by immunization of the purified recombinant protein.The level of Cu/Zn SOD expression in B.abortus S19 was analyzed by SDS-PAGE.Our data indicated that Cu/Zn SOD which expressed through the inducible protein expression system pBBR-lacPtrc-sod had high expression level and good specificity.In conclusion,Cu/Zn SOD was overexpressed successfully in B.abortus S19 in this research.Additionally,these results also implied that our expression system can be applied in antigen modification of B.abortus S19.

关 键 词：布鲁菌 S19疫苗株 铜锌超氧化物歧化酶 

分 类 号：S852.61[农业科学—基础兽医学] R535[农业科学—兽医学]