表达PEDV S蛋白重组PRV的构建与生物学特性评价  

Construction and biological characteristics evaluation of recombinant PRV expressing S protein of PEDV

作　　者：侯晓璇 穆永 王同燕 颜世君 王孟月 谭菲菲 田克恭 HOU Xiaoxuan;MU Yong;WANG Tongyan;YAN Shijun;WANG Mengyue;TAN Feifei;TIAN Kegong(College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China;National Research Center for Veterinary Medicine,Luoyang,Henan 471000,China;PULIKE Biological Engineering Co.,Ltd.,Luoyang,Henan 471000,China)

机构地区：[1]河南农业大学动物医学学院,河南郑州450046 [2]国家兽用药品工程技术研究中心,河南洛阳471000 [3]普莱柯生物工程股份有限公司,河南洛阳471000

出　　处：《中国兽医学报》2024年第4期657-663,共7页Chinese Journal of Veterinary Science

基　　金：郑洛新自创区创新引领型产业集群专项资助项目(201200211200)。

摘　　要：将猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)流行毒株的S胞外区基因克隆至含有gG基因上、下游同源臂以及抗性筛选基因的转移质粒pUC-ABK中,构建供体质粒pUC-ABK-S,获得线性同源重组片段AS-K-B。利用Red同源重组技术将线性同源重组片段电转化至含有pPRVBac-GFPTK-gE-gI-11k-28k-的感受态中获得重组pPRVBac-S,最后拯救获得重组病毒rPRV-S。PCR及IFA鉴定结果显示,S胞外区基因成功转录且表达,生物学特性评价结果表明重组病毒能够稳定遗传,为研制猪流行性腹泻病毒活载体疫苗提供了思路。In order to construct a recombinant pseudorabies virus expressing S protein of porcine epidemic diarrhea virus,the S extracellular region gene of porcine epidemic diarrhea virus epidemic strain was cloned into the transfer plasmid pUC-ABK containing the upstream and downstream homologous arms of gG gene and the resistance screening gene to construct the donor plasmid pUC-ABK-S,and the linear homologous recombination fragment A-S-K-B was obtained.The linear homologous recombinant fragment was electrotransformed into the competent cells containing pPRVBac-GFPTK-gE-gl-11k-28k-to obtain recombinant pPRVBac-S by Red homologous recombination,and finally,the recombinant virus rPRV-S was rescued.PCR and IFA identification results showed that S extracellular region gene was successfully transcribed and expressed,the results of biological characteristics evaluation showed that the recombinant virus could be stably inherited,which provided an experimental basis for the development of porcine epidemic diarrhea virus live vector vaccine.

关 键 词：PEDV S蛋白 RED同源重组 重组PRV 

分 类 号：S852.65[农业科学—基础兽医学]