A型塞内卡病毒VP1蛋白的原核表达及多克隆抗体制备  

Prokaryotic Expression of VP1 Protein of Senecavirus A and Preparation of Its Polyclonal Antibodies

作　　者：秦文珍 李挺 赵欣[1,2] 董苏洁 翟焕杰 叶晨倩 叶曼青 童武 郑浩[2] 于海[2] 单同领[2] 童光志[2] 兰道亮[1] 孔宁[2] QIN Wenzhen;LI Ting;ZHAO Xin;DONG Sujie;ZHAI Huanjie;YE Chenqian;YE Manqing;TONG Wu;ZHENG Hao;YU Hai;SHAN Tongling;TONG Guangzhi;LAN Daoliang;KONG Ning(College of Animal&Veterinary Sciences,Southwest Minzu University,Chengdu 610041,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China)

机构地区：[1]西南民族大学畜牧兽医学院,成都610041 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2024年第2期195-200,共6页Chinese Journal of Animal Infectious Diseases

基　　金：西南民族大学中央高校基本科研业务费专项资金项目(2020YYXS70)。

摘　　要：A型塞内卡病毒(SVA)是一种新兴的猪病原体,可引起母猪水泡样病变和仔猪急性死亡。本研究将SVA的VP1基因分别克隆到原核表达载体pCold-I和pCold-TF,并将测序正确的重组质粒VP1-pCold-I、VP1-pCold-TF转化BL21(DE3)大肠杆菌中,诱导表达重组蛋白。结果显示:VP1-pCold-I表达的目的蛋白在沉淀(包涵体)中,VP1-pCold-TF表达的蛋白为可溶性蛋白。分别纯化上述表达的蛋白,并免疫BALB/c小鼠,经四次免疫后得到多克隆抗体。经Western blot和间接免疫荧光(IFA)鉴定,制备的多克隆抗体具有良好的Western blot效价和IFA效价,为相关基础研究和应用研究提供了工具。Senecavirus A(SVA)is an emerging swine pathogen that can cause vesicular lesions in sows and acute death of piglets.In this study,the VP1 gene of SVA was cloned into the prokaryotic expression vectors pCold-I and pCold-TF and then the recombinant plasmids VP1-pCold-I and VP1-pCold-TF were verified and transformed into E.coli BL21(DE3)to induce the expression of recombinant protein.The results showed that VP1-pCold-I existed in the precipitates(inclusion bodies)and VP1-pCold-TF was a soluble protein.The purified proteins were used to immunize BALB/c mice.The polyclonal antibodies were obtained after immunization four times and detected in Western blotting and indirect immunofluorescence assay(IFA).The prepared polyclonal antibodies had good tiers in Western blot and IFA,which provided a tool for relevant basic and applied research.

关 键 词：A型塞内卡病毒 VP1 原核表达 多克隆抗体 

分 类 号：S852.65[农业科学—基础兽医学]