中国动物检疫2024，Vol.41Issue(4)：96-102,7.DOI:10.3969/j.issn.1005-944X.2024.04.018
A型塞内卡病毒VP3蛋白的原核表达及多克隆抗体制备
Prokaryotic Expression of the VP3 Protein of Senecavirus A and Preparation of Its Polyclonal Antibodies
摘要
Abstract
In order to prepare polyclonal antibodies against VP3 protein of Senecavirus A(SVA),a recombinant expression plasmid named as pET-SVA-VP3 was constructed through cloning SVA VP3 gene into a prokaryotic expression vector known as pET-28a using homologous recombination technology,and transformed into Escherichia coli competent cell BL21(DE3)to express VP3 protein by IPTG induction at a concentration of 1 mmol/L,and the expressed protein was purified,then the purified VP3 recombinant protein was injected into New Zealand white rabbits via subcutaneous multi-point to prepare polyclonal antibodies,followed by evaluation on its titer,reactivity and specificity by indirect ELISA,indirect immunofluorescence assay(IFA)and Western blot.The results showed that the VP3 recombinant protein was expressed in a form of inclusion body with a molecular mass of about 36 kDa;the prepared antibodies could specifically react with VP3 recombinant protein and SVA,and the titer evaluated by ELISA reached up to above 1:10 000,and those by Western blot and IFA were both 1:5 000.In conclusion,SVA VP3 polyclonal antibodies were successfully prepared,which were with high titer and specificity.The study laid a foundation for future researches on the pathogenic mechanism and detection of SVA.关键词
A型塞内卡病毒/VP3蛋白/原核表达/多克隆抗体/效价/特异性Key words
SVA/VP3 protein/prokaryotic expression/polyclonal antibody/titer/specificity分类
畜牧业引用本文复制引用
林铱婷,田文霞,任建乐,姬康,谭姗姗,晋怡,宋若琪,马瑞一,王颖,牛胜,赵宇军..A型塞内卡病毒VP3蛋白的原核表达及多克隆抗体制备[J].中国动物检疫,2024,41(4):96-102,7.基金项目
山西省科技创新人才团队专项(202204051001022) （202204051001022）
山西省优秀博士来晋奖励基金项目(SXBYKY2021036) （SXBYKY2021036）
山西农业大学博士科研启动基金项目(2020BQ59) （2020BQ59）
