A型塞内卡病毒的病毒样颗粒的制备及其免疫原性分析  

Generation and immunogenicity evaluation of Senecavirus A virus-like particles

作　　者：伍春平 茹毅 田宏[1] 马坤 郝荣增 李亚军 罗俊聪 石正旺 刘华南[1] 左志 郑海学[1] Chunping Wu;Yi Ru;Hong Tian;Kun Ma;Rongzeng Hao;Yajun Li;Juncong Luo;Zhengwang Shi;Huanan Liu;Zhi Zuo;Haixue Zheng(State Key Laboratory of Veterinary Etiological Biology,OIE/National Foot-and-Mouth Disease Reference Laboratory,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,Gansu,China)

机构地区：[1]中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室OIE/国家口蹄疫参考实验室,甘肃兰州730046

出　　处：《生物工程学报》2021年第9期3211-3220,共10页Chinese Journal of Biotechnology

基　　金：甘肃省科技重大专项(No.19ZDNA001);洛阳市科技重大专项(No.1901029A)资助。

摘　　要：为了研制A型塞内卡病毒(Senecavirus A,SVA)的病毒样颗粒(Virus-like particles,VLPs)疫苗,以SVA田间流行毒株CH-FJ-2017结构蛋白基因序列为研究对象,构建了能够同时表达SVA的3种结构蛋白VP0、VP1和VP3的单个原核重组表达质粒pET28a-SVA-VP031。通过大肠杆菌Escherichia coli表达、亲和层析纯化和体外自组装,获得SVA VLPs。透射电子显微镜鉴定显示,SVA的3种结构蛋白在体外能够自组装成直径约25–30 nm的VLPs,并且动物免疫试验结果表明,该VLPs能够有效刺激豚鼠产生高水平的抗原特异性中和抗体。上述研究结果为SVA VLPs疫苗的研制奠定了基础。To develop Senecavirus A(SVA) virus-like particles(VLPs), a recombinant prokaryotic expression plasmid pET28 a-SVA-VP031 was constructed to co-express SVA structural proteins VP0, VP3 and VP1, according to the genomic sequence of the field isolate CH-FJ-2017 after the recombinant proteins were expressed in E.coli system, and purified by Ni+ ion chromatographic method. The SVA VLPs self-assemble with a high yield in vitro buffer. A typical VLPs with an average diameter of 25–30 nm which is similar to native virions by using TEM detection. Animals immunized by SVA VLPs shown that the VLPs induced high titers neutralizing antibodies in Guinea pigs. This study indicated that the VLPs produced with co-expressing SVA structural proteins VP0, VP3 and VP1 in prokaryotic system is a promising candidate and laid an important foundation for the development of a novel SVA VLPs vaccine.

关 键 词：A型塞内卡病毒 病毒样颗粒 结构蛋白 免疫原性 

分 类 号：S852.65[农业科学—基础兽医学]