水痘-带状疱疹病毒ORF43蛋白的单克隆抗体制备及初步研究  

Preparation and Preliminary Study of Monoclonal Antibodies Against Varicella-zoster Virus ORF43

作　　者：翟兵可 潘德全 付文锟 蔡雨泽 王玮 赵欢[1,2] 阙玉琼 程通[1,2] 夏宁邵[1,2] ZHAI Bingke;PAN Dequan;FU Wenkun;CAI Yuze;WANG Wei;ZHAO Huan;QUE Yuqiong;CHENG Tong;XIA Ningshao(National Institute of Diagnostics and Vaccine Development in Infectious Diseases,School of Life Science,Xiamen University,Xiamen,361102,China;State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics,School of Public Health,Xiamen University,Xiamen,361102,China)

机构地区：[1]国家传染病诊断试剂与疫苗工程技术研究中心,厦门大学生命科学学院,厦门361102 [2]分子疫苗学和分子诊断学国家重点实验室,厦门大学公共卫生学院,厦门361102

出　　处：《病毒学报》2021年第6期1302-1309,共8页Chinese Journal of Virology

基　　金：国家自然科学基金(项目号:81871648),题目:水痘-带状疱疹病毒的衣壳结构与衣壳蛋白相互作用机制研究。

摘　　要：水痘-带状疱疹病毒(Varicella-zoster virus,VZV)是引起水痘和带状疱疹这两种临床表现不同病症的共同致病原,其基因组中ORF43是VZV在宿主细胞中复制的必需基因,但目前尚无针对VZV ORF43编码蛋白性质与功能的研究报道。本研究目的是制备抗VZV ORF43单克隆抗体,以初步研究该蛋白在细胞内的表达与分布情况。本研究构建了VZV ORF43蛋白的原核表达质粒并在大肠杆菌中进行了该蛋白的表达,纯化蛋白免疫小鼠后,使用杂交瘤技术及克隆化筛选,获得一株特异性强、反应性好的抗VZV ORF43单克隆抗体2D3。本研究使用该抗体鉴定出VZV ORF43蛋白在质粒转染细胞与病毒感染细胞中表达的分子量大小均约为70kD,但分别呈现出细胞质和细胞核的不同亚细胞定位。以上工作为后续进一步探究该蛋白在VZV感染与致病过程中的作用奠定了基础。Varicella zoster virus(VZV)is the common causative agent of chickenpox and shingles,which show different clinical manifestations. Open reading frame(ORF)43 is an essential gene for VZV replication in host cells,but the nature and function of its encoded viral protein has not been reported. The purpose of this study is to prepare monoclonal antibodies(mAbs) against VZV ORF43 protein(pORF43) and determine the expression and subcellular distribution of this protein in cells. A prokaryotic expression plasmid of VZV pORF43 was constructed,and recombinant pORF43 was expressed and purified from Escherichia coli. After immunizing mice with the purified protein,a highly specific and sensitive anti-VZV pORF43 mAb(2 D3)was obtained by the hybridoma method and clonal screening. Using 2 D3,VZV pORF43 was identified to be a protein of approximately 70 kD. Notably,this protein was predominantly localized in the cytoplasm and nucleus in plasmid - transfected cells and VZV - infected cells,respectively. These data lay a groundwork for further research on the role of pORF43 in VZV infection and pathogenesis.

关 键 词：水痘-带状疱疹病毒(VZV) ORF43 单克隆抗体 

分 类 号：R373.9[医药卫生—病原生物学]