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 ex-pressing S protein of PEDV
侯晓璇 1穆永 1王同燕 2颜世君 3王孟月 3谭菲菲 3田克恭2
作者信息
- 1. 河南农业大学动物医学学院,河南 郑州 450046
- 2. 河南农业大学动物医学学院,河南 郑州 450046;国家兽用药品工程技术研究中心,河南 洛阳 471000;普莱柯生物工程股份有限公司,河南洛阳 471000
- 3. 国家兽用药品工程技术研究中心,河南 洛阳 471000;普莱柯生物工程股份有限公司,河南洛阳 471000
- 折叠
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A-S-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.
Abstract
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 pro-vided an experimental basis for the development of porcine epidemic diarrhea virus live vector vac-cine.
关键词
PEDV/S蛋白/Red同源重组/重组PRVKey words
PEDV/S protein/Red homologous recombination/recombinant PRV引用本文复制引用
基金项目
郑洛新自创区创新引领型产业集群专项(201200211200)
出版年
2024
NETL
NSTL
万方数据