重组牛源荚膜A型多杀性巴氏杆菌脂蛋白E的免疫保护性研究  

Protective immunity induced by recombinant lipoprotein E(PlpE) of bovine Pasteurella multocida type A

作　　者：牛思博[1] 姜志刚[1] 德艳艳[1] 于力[1] 

机构地区：[1]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/大动物病研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2013年第5期401-404,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省科技攻关项目(GA12B301)

摘　　要：为鉴定多杀性巴氏杆菌脂蛋白E的免疫保护性,本研究从牛源荚膜A型多杀性巴氏杆菌PM-HLJ株的基因组DNA中扩增出大小为1 011 bp的脂蛋白E编码基因(PlpE),经序列测定显示,其氨基酸序列与A型多杀性巴氏杆菌参考株PlpE序列一致性为95.83%。将PM-HLJ的PlpE基因克隆于表达质粒pET-30a(+)并诱导表达,将表达的重组脂蛋白E纯化后经腹腔免疫小鼠,间接ELISA检测结果显示各免疫组均产生了针对重组脂蛋白E的IgG抗体,免疫保护试验显示,当小鼠免疫剂量为20μg、30μg、40μg和60μg时,保护率分别达到20%、40%、80%和100%。结果表明,牛源荚膜A型多杀性巴氏杆菌重组脂蛋白E具有良好的免疫保护作用。To evaluate the immunoprotection of Pasteurella multocida lipoprotein E （PlpE）, a 1,011 bp PlpE encoding gene from bovine P.multocida type A strain PM-HLJ was amplified by PCR. The sequence analysis showed the amino acid sequence of the PIpE from PM-HLJ strain exhibited 95.83% identity with that of reference strain. In addition, the PlpE gene was cloned into pET-30a（＋） for expression in E.coli, and the expression of soluble recombinant PIpE （rPlpE） was induced with 0.2 mM IPTG at 16 ~C. Furthermore, rPlpE was able to produce protection rate of 20%, 40%, 80% or 100% against the challenging with 3 MLD of P.multocida type A strain PM-HLJ in mice when immunized with purified rPlpE at dose of 20 μg, 30 μg, 40 μg and 60 μg, respectively. The results demonstrated the rPlpE was capable of inducing efficient protective immune response in mice and had potential to be a desirable immunogen for stimulating protective immunity against P.multocida type A in cattle.

关 键 词：牛源荚膜A型多杀性巴氏杆菌 重组脂蛋白E 免疫保护 

分 类 号：S852.61[农业科学—基础兽医学]