HCV结构蛋白在昆虫细胞中的表达及病毒样颗粒的组装  

Expression of HCV Structural Proteins and Self-assembly of HCV-like Particles in Insect Cells

作　　者：王瑞青[1] 梁昌镛[1] 焦成松[2] 宋建华[1] 陈新文[1] 

机构地区：[1]中国科学院武汉病毒研究所分子病毒学重点实验室 [2]中国人民解放军广州军区武汉总医院,湖北武汉430071

出　　处：《中国病毒学》2005年第1期5-10,共6页Virologica Sinica

基　　金：湖北省自然科学基金(2003ABA157);国家自然科学基金杰出青年基金(30325002)

摘　　要：本文利用Bac to Bac杆状病毒表达系统构建了含有丙型肝炎病毒 (Hepatitis C Virus,HCV)结构蛋白编码基因的重组杆状病毒vAcHCVsp1,并获得了HCV结构蛋白在昆虫细胞Sf21中的表达。HCV mRNA转录和蛋白质表达时相分析表明,感染后16 h HCV结构蛋白编码基因开始转录,72 h达最高峰;蛋白质表达则是在感染后48h开始,72 h达到高峰。电镜观察表明 vAcHCVsp1 感染的 Sf21 细胞 96 h时在细胞质中可见很多空泡,空泡中可见50nm的球形颗粒,为HCV结构蛋白组装的病毒样颗粒。In this work, a recombinant baculovirus containing Hepatitis C virus (HCV) structural protein coding sequence was constructed by Bac-to-Bac recombinant baculovirus expression system and it was proved to be able to produce HCV structural proteins directly in insect cells. RT-PCR assay showed that the transcription of the coding sequence began 16h post infection (hpi) and reached maximal level at 72 hpi. Analysis of insect cell lysates by SDS-PAGE and immuno-blotting with monoclonal antibody against Core revealed that the expression of the structural proteins began at 48 hpi and reached maximal level also at 72hpi. In insect cells, HCV structural proteins assembled into spherical particles 50nm in diameter in large cytoplasmic vacuoles.

关 键 词：丙型肝炎病毒 结构蛋白 表达 病毒样颗粒 Sf21细胞 

分 类 号：R346[医药卫生—基础医学]