摘要
为了探究非洲猪瘟病毒(ASFV)A137R蛋白的表达动力学与亚细胞定位,本研究构建了原核表达质粒pGEX-6p-1-A137R,表达与纯化了A137R蛋白,并制备了针对该蛋白的多克隆抗体。经间接免疫荧光试验(IFA)分析,抗体效价达1∶25600;Western-blotting和IFA证实,该多克隆抗体具有良好的反应性和特异性。利用该抗体分析了ASFV感染原代猪肺泡巨噬细胞中A137R蛋白的表达动力学及其亚细胞定位,发现A137R蛋白为晚期蛋白,主要定位于细胞质,且与ASFV的结构蛋白p72共定位。本研究为进一步解析ASFV A137R蛋白的功能奠定了基础。
To identify the expression kinetics and subcellular localization of the A137R protein,the protein was expressed in E.coli and the polyclonal antibodies against the protein were prepared.The polyclonal antibodies,with a titer of 1∶25600 showed a great reactivity and specificity,as confirmed by Westernblotting and indirect immunofluorescent assay(IFA).The expression kinetics and subcellular localization of the A137R protein in the ASFV-infected primary porcine alveolar macrophages,indicated that the A137R protein is a late-expressed protein,mainly localizes in the cytoplasm and colocalizes with the ASFV structural protein p72.The study provides the reference for research on the functions of the ASFV A137R protein.
作者
孙茂文
周萍萍
王涛
李永锋
潘力
韩玉
孙元
杨玉莹
仇华吉
SUN Mao-wen;ZHOU Ping-ping;WANG Tao;LI Yong-feng;PAN Li;HAN Yu;SUN Yuan;YANG Yu-ying;QIU Hua-ji(College of Animal Sciences,Yangtze University,Jingzhou 434023,China;State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2021年第12期1525-1532,共8页
Chinese Veterinary Science
基金
国家自然科学基金面上项目(32072854)
国家自然科学基金联合基金项目(U20A2060)。
关键词
非洲猪瘟病毒
A137R蛋白
原核表达
多克隆抗体
African swine fever virus
A137R protein
prokaryotic expression
polyclonal antibodies
