牛多杀性巴氏杆菌znuA基因的原核表达及其免疫保护作用的研究  

Prokaryotic expression of znuA of bovine Pasteurella multocida and its immunoprotection in mice

作　　者：嵇少泽 高云航[1] 勾长龙 张喜庆[1] 田佳琪[1] 李晨 王思月 JI Shao-ze;GAO Yun-hang;GOU Chang-long;ZHANG xi-qing;TIAN Jia-qi;LI Chen;WANG Si-yue(Department of Preventive Veterinary Medicine,川in Agricultural University,Changchun 130118,China)

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118

出　　处：《中国预防兽医学报》2019年第2期184-187,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：肉牛产业技术体系废弃物处理与环境控制岗位(CARS-38);畜禽重大疫病防控与高效安全养殖综合技术研发(2016YFD0501400);畜禽养殖固体废弃物无害化处理及资源化利用技术研究(20170101073JC);养殖场固体废弃物处理功能性菌剂研发及资源化利用技术研究(JJKH20180643KJ)

摘　　要：为研究牛多杀性巴氏杆菌(P.multocida)高亲和力锌吸收蛋白znuA的免疫学活性及其免疫保护作用,本研究利用PCR方法扩增了P.multocida znuA基因,构建表达载体pET-28a-znuA,将其转化E.coli BL21后经诱导表达。表达产物经SDS-PAGE和western blot分析显示,重组蛋白约40 ku;以重组znuA蛋白免疫小鼠后用P.multocida菌株Y-1攻毒,结果显示重组znuA蛋白对免疫组小鼠保护率为60%,表明其具有免疫保护作用。本研究首次在原核系统中表达了P.multocida znuA蛋白,且验证了其免疫保护力,为深入探究znuA基因在P.multocida致病过程中的作用及其亚单位疫苗的开发奠定了基础。To study the immunoprotection of high-affinity zinc uptake system protein znuA against in mice. The target gene,znuA, was amplified by PCR from Pasteurella multocida of Bovine and cloned into the pET-28a vector. The constructed recombinant plasmid pET-28a-znuA was transformed to E.coli BL21(DE3)for expression under induction of IPTG. SDS-PAGE and western blot analysis showed that the expressed recombinant znuA was about 40 ku. The mice were immunized intramuscularly with purified recombinant znuA, and challenged with the Pasteurella multocida Y-1, result showed that the recombinant znuA provided a 60% protection for the immunized mice. All results indicated that the target protein can be over expressed in E.coli BL21, which would induced protective immune response in mice, indicating its potential as an antigen of Pasteurella multocida protein vaccine.

关 键 词：多杀性巴氏杆菌 znuA 保护性免疫 

分 类 号：S852.61[农业科学—基础兽医学]