复制缺陷型痘苗病毒天坛株复制缺陷机制初步探究  

Preliminary exploration of replication-defective mechanism of highly attenuated NTV strain ofvaccinia virus Tiantan

作　　者：黄盼盼 赵莉[1] 任皎[1] 赵颖 阮力[1] 谭文杰[1] 田厚文[1] Huang Panpan;Zhao Li;Ren Jiao;Zhao Ying;Ruan Li;Tan Wenjie;Tian Houwen(National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China)

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京102206

出　　处：《中华实验和临床病毒学杂志》2018年第2期119-123,共5页Chinese Journal of Experimental and Clinical Virology

基　　金：国家科技重大专项（2013ZX10004001,2013ZX10004101）;国家科技重点研发项目（2016YFC1200900）

摘　　要：目的检测痘苗病毒早期、晚期蛋白表达水平，对复制缺陷型痘苗病毒天坛株（NTV）复制缺陷机制进行初步探究。方法构建原核表达载体表达并纯化与痘苗病毒复制密切相关的早蛋白E3和晚期蛋白F17，免疫小鼠制备相应鼠多抗血清，用其对NTV病毒在非允许细胞中相应蛋白的表达水平进行检测。结果在大肠埃希菌中分别表达、纯化了痘苗病毒E3和F17蛋白，并制备获得了相应的鼠多抗血清；Westernblot鉴定在非允许细胞Hela中复制缺陷型痘苗病毒NTV的早期蛋白E3及晚期蛋白A27能正常表达，但晚期蛋白F17表达受到明显抑制。结论F17蛋白表达受限可能是NTV复制缺陷的机制之一。Objective To detect the expression level of early and late protein of vaccinia virus and to preliminarily explore replication-defective mechanism of highly attenuated NTV strain of vaccinia virus Tiantan. Methods We constructed prokaryotie expression vector, expressed and purified homologous early protein E3 and late protein A27 closely related to replication and prepared mouse polyclonal antiserum by immunizing mice with homologous proteins. Early and late protein expression levels of NTV were detected. Results We have expressed and purified vaccinia virus proteins respectively in E. coli expression system and prepared homologous mouse polyclonal antiserum. Early protein E3 and late protein A27 could be highly efficient expression in NTV infected non-permissive Hela cells, while expression of late protein F17 was blocked detected by Western blot. Conclusions The expression limitation of late protein F17 may be an explanation for the replication-defective mechanism of NTV.

关 键 词：复制缺陷型痘苗病毒 痘苗病毒载体 NTV 复制缺陷机制 

分 类 号：R392[医药卫生—免疫学]