HSV-1HF株扩增子载体的构建及其在不同HSV血清型间的通用性研究  

Construction of the HSV-1 Strain HF Amplicon and Study on Its Unversal Function between Different HSV Serotypes

作　　者：宋波[1] 刘新静[1] 韩志强[1] 赵璐[1] 王青志[1] 卢甲盟[1] 许予明[1] 

机构地区：[1]郑州大学第一附属医院神经内科,郑州450052

出　　处：《病毒学报》2011年第5期409-415,共7页Chinese Journal of Virology

基　　金：河南省卫生厅医学科技攻关项目(200801003);河南省卫生厅医学科技攻关项目(200702005);河南省教育厅河南省基础与前沿技术研究计划项目(082300450270);河南省教育厅自然科学研究计划项目(2009A320045)

摘　　要：旨在构建HSV-1HF株的扩增子载体,研究其在不同血清型HSV辅助下的包装通用性。经酶切HF株的BAC-HSV-1,获得oriS和pac元件并测序。以pSilencer2.0-U6为骨架,以DsRed为报告基因构建HSV-1HF株的扩增子载体,利用脂质体2000转染扩增子载体至Vero细胞,分别应用HSV-1HF株和HSV-2HG52辅助HSV-1扩增子载体进行包装,待产生细胞病变效应后取上清,再次感染Vero细胞,观察Vero细胞内红色荧光蛋白表达情况。本研究首次构建了HSV-1HF株的扩增子载体,鉴定了HSV-1HF株oriS和pac元件,HSV-1HF株扩增子载体可以被HSV-1HF株和HSV-2HG52株包装并扩增。The study aimed to construct the amplicon vector of HSV-1 strain HF and explore its universal package function between different serotypes of HSV. OriS and pac elements were obtained by enzyme di- gestion from the Plasmid BAC-HSV-1 strain HF and sequenced. With red fluorescence （DsRed） as a reporter gene, the amplicon vector of HSV-1 strain HF was constructed based on pSilencer2.0-U6. The amplicon vector was transfected into Vero cells by lipofectamine 2000, then packaged by HSVq strain HF and HSV-2 strain HG52 as helper virus separately. The supernatant was collected after cytopathic effect. Red fluorescence was observed in Veto cells reinfected by the supernatant. In this study,the amplicon vector of HSV-1 strain HF was successfully constructed and it could be packaged by HSV-1 strain HF and HSV-2 strainHG52.

关 键 词：单纯疱疹病毒I型 HSV-1 HF株 HSV 2 HG52株 复制起始点 包装信号 扩增子载体 

分 类 号：R373.11[医药卫生—病原生物学]