乙型脑炎病毒部分NS1基因假病毒的构建  

Construction of pseudoviruses containing a partial NS1 gene of Japanese encephalitis virus

作　　者：龙彩韵 王瑞晨 姚晓慧 武婕慧 付士红[2] 李樊 殷启凯 崔倩倩 许松涛[2] 聂凯[2] 王环宇[2] LONG Cai-yun;WANG Rui-chen;YAO Xiao-hui;WU Jie-hui;FU Shi-hong;LI Fan;YIN Qi-kai;CUI Qian-qian;XU Song-tao;NIE Kai;WANG Huan-yu(Medical School,Anhui University of Science and Technology,Huainan 232001,China;National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China)

机构地区：[1]安徽理工大学医学院,淮南232001 [2]中国疾病预防控制中心病毒病预防控制所,北京102206

出　　处：《中国人兽共患病学报》2024年第3期231-235,242,共6页Chinese Journal of Zoonoses

基　　金：国家重点研发计划(No.2020YFE0205700,No.2022YFC2302700)。

摘　　要：目的构建耐核酸酶的含乙型脑炎病毒(Japanese encephalitis virus,JEV)部分非结构蛋白1(Non-structural protein 1,NS1)基因的假病毒,为JEV的实验室核酸检测提供质控品,提高检测质量。方法将JEV部分NS1基因、蚊18S RNA基因和人GAPDH基因片段次序连接克隆到该质粒上,构建重组载体pET-MS2-JEV;转化大肠杆菌后表达纯化假病毒,并对其进行耐受性和稳定性评估。结果构建了重组载体pET-MS2-JEV,表达获得的假病毒具有耐核酸酶、耐物理化学因子的特性,可在4℃和-20℃条件下稳定保存。结论本研究获得了稳定性好的含JEV部分NS1基因的假病毒,可作为JEV实验室核酸检测的质控品。Construction of pseudoviruses containing a partial non-structural protein 1(NS1)gene of Japanese encephalitis virus(JEV)with nuclease resistance for application as quality control reagents in laboratory assay.Firstly,the plasmid pET-32a-MS2 underwent optimization and modification.Secondary,fragments of the partial NS1 gene of the JEV,along with internal reference genes from mosquitoes and humans(mosquito 18S RNA and human GAPDH),were inserted into this plasmid,resulting in the construction of the recombinant plasmid pET-MS2-JEV.Subsequent steps involved the expression and purification of pseudoviruses.The construction of the recombinant plasmid pET-MS2-JEV yielded pseudoviruses,which obtained resilience against nucleases and various physical and chemical factors.These pseudoviruses exhibited stability when stored at various temperature conditions,including 4℃and-20℃.This study accomplished the production of pseudoviruses characterized by outstanding stability,thus serving as reliable quality control reagents for JEV laboratory assays.

关 键 词：乙型脑炎病毒 MS2噬菌体 假病毒 

分 类 号：R373.9[医药卫生—病原生物学]