牛病毒性腹泻病毒NS5B蛋白的截短表达及多克隆抗体制备  

Prokaryotic Expression of Truncated BVDV NS5B Protein and Preparation of Polyclonal Antibodies

作　　者：秦春雪 夏国建 何成强[2] QIN Chunxue;XIA Guojian;HE Chengqiang(Key Laboratory of Biopharmaceuticals of Shandong Province,New Drug Center of Shandong Academy of Pharmaceutical Sciences,Jinan 250014,China;Shandong Key Laboratory of Animal Resistance,College of Life Science,Shandong Normal University,Jinan 250014,China)

机构地区：[1]山东省药学科学院新药评价中心山东省生物制药重点实验室,济南250014 [2]山东师范大学生命科学学院山东省动物抗性重点实验室,济南250014

出　　处：《中国动物传染病学报》2024年第4期167-172,共6页Chinese Journal of Animal Infectious Diseases

基　　金：山东省重点研发项目(2017GNC10125,2019GSF107020)。

摘　　要：本研究旨在克隆牛病毒性腹泻病毒(BVDV)的非结构蛋白NS5B截短基因,采用原核表达系统表达获得截短蛋白,并进一步制备其多克隆抗体。构建的原核表达质粒pET-28a-NS5B转化至BL21感受态细胞,经IPTG诱导后,发现NS5B蛋白以包涵体为主。纯化蛋白免疫小鼠制备的多克隆抗体具有良好的反应性,能够特异性识别真核表达的NS5B蛋白。本研究中NS5B蛋白的截短表达及其多克隆抗体的制备为进一步研究NS5B蛋白的生物学功能,以及建立BVDV鉴别诊断的血清学检测方法奠定了基础。The purpose of this study was to clone the gene fragment of the non-structural protein NS5B of Bovine viral diarrhea virus(BVDV)for prokaryotic expression of the truncated protein and prepare its polyclonal antibodies.The constructed plasmid pET-28a-NS5B was transformed into BL21 competent cells.The truncated NS5B protein was expressed with induction of IPTG and mainly released as inclusion bodies.The polyclonal antibodies prepared by immunizing mice had good reactivity and specifically recognized the eukaryotic expressed NS5B protein.The expression of truncated NS5B protein and preparation of polyclonal antibodies in this study laid the foundation for further studying the biological function of NS5B protein and developing a serological method for differential diagnosis of BVDV.

关 键 词：牛病毒性腹泻病毒 NS5B蛋白 原核表达 多克隆抗体 

分 类 号：S852.65[农业科学—基础兽医学]