猪繁殖与呼吸综合征病毒非结构蛋白7多克隆抗体的制备及鉴定  

Preparation and Identification of Polyclonal Antibodies against Nonstructural Protein 7 of Porcine Reproductive and Respiratory Syndrome Virus

作　　者：李华玮[1] 李莎 姬向波 刘昆 杨中元 侯文静[1] 马辉[1] 赵绪永[1] LI Hua-wei;LI Sha;JI Xiang-bo;LIU Kun;YANG Zhong-yuan;HOU Wen-jing;MA Hui;ZHAO Xu-yong(Henan University of Animal Husbandry and Economy,Zhengzhou,Henan,450046,China;Henan Key Laboratory of Innovation and Utilization of Unconventional Feed Resources,Zhengzhou,Henan,450046,China;Henan Agricultural University,Zhengzhou,Henan,450046,China)

机构地区：[1]河南牧业经济学院,河南郑州450046 [2]河南省非常规饲料资源创新利用重点实验室,河南郑州450046 [3]河南农业大学,河南郑州450046

出　　处：《动物医学进展》2023年第6期15-21,共7页Progress In Veterinary Medicine

基　　金：河南省科技研发计划联合基金项目(应用攻关类)(222103810022);河南省科技攻关计划项目(232102110104;202102110245);河南牧业经济学院预防兽医重点学科项目(XJXK202202)。

摘　　要：制备猪繁殖与呼吸综合征病毒(PRRSV)非结构蛋白7(nsp7)的多克隆抗体。根据NCBI上公布的PRRSV nsp7序列,合成基因后克隆至原核表达载体pET28a,对重组菌进行诱导,获得表达并纯化PRRSV nsp7重组蛋白。采用皮下多点注射的方式对新西兰白兔进行免疫接种制备兔多克隆血清,分别使用ELISA、IFA、Western blot等方法验证该多克隆血清与PRRSV以及nsp7重组蛋白的免疫反应性。间接ELISA检测结果显示,制备的多抗血清可以特异性识别原核表达的nsp7,IFA检测结果证实制备的nsp7多抗能特异性结合PRRSV,Western blot结果证实多抗血清能特异性结合PRRSV以及纯化的重组蛋白nsp7。以上结果表明,成功制备了PRRSV nsp7的多克隆抗体。Preparation of polyclonal antibodies to non-structural protein 7(nsp7)of porcine reproductive and respiratory syndrome virus(PRRSV)was conducted.The PRRSV nsp7 recombinant protein was obtained and purified by cloning the gene into the pET28a vector based on the PRRSV nsp7 sequence published in the NCBI,and inducing expression in recombinant bacteria.The rabbits were immunized by subcutaneous multi-point injections,and the rabbit polyclonal antibody serum was prepared.The immunoreactivity of the polyclonal antibody serum with PRRSV and nsp7 recombinant protein was verified by ELISA,IFA and Western blot methods,respectively.The results of indirect ELISA showed that the prepared polyclonal antibodies specifically recognized the prokaryotic expression of nsp7,IFA confirmed that the prepared nsp7 polyclonal antibodies specifically bound PRRSV,and Western blot confirmed that the polyclonal antibodies specifically bound PRRSV and the purified recombinant protein nsp7.The polyclonal antibodies to PRRSV nsp7 were successfully prepared.

关 键 词：猪繁殖与呼吸综合征病毒 非结构蛋白7 多克隆抗体 

分 类 号：S852.659.6[农业科学—基础兽医学] S852.651[农业科学—兽医学]