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
秦文珍 1李挺 1赵欣 1董苏洁 2翟焕杰 2叶晨倩 2叶曼青 2童武 2郑浩 2于海 2单同领 2童光志 2兰道亮 3孔宁2
作者信息
- 1. 西南民族大学畜牧兽医学院,成都 610041;中国农业科学院上海兽医研究所,上海 200241
- 2. 中国农业科学院上海兽医研究所,上海 200241
- 3. 西南民族大学畜牧兽医学院,成都 610041
- 折叠
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.
Abstract
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Key words
Senecavirus A/VP1/prokaryotic expression/polyclonal antibody引用本文复制引用
基金项目
中央高校基本科研业务费专项西南民族大学项目(2020YYXS70)
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据