中国株丙型肝炎病毒(HCV)结构区蛋白在昆虫细胞中的表达及加工  

THE EXPRESSION AND PROCESSING OF STRUCTURAL POLYPROTEIN OF CHINESE HEPATITIS C VIRUS EXPRESSED IN INSECT CELLS

作　　者：苗季[1] 谭文杰[1] 丛旭[1] 丛郁[1] 陈刚[1] 毕胜利[1] 詹美云[1] 

机构地区：[1]中国预防医学科学院病毒学研究所

出　　处：《病毒学报》1998年第4期289-295,共7页Chinese Journal of Virology

摘　　要：利用杆状病毒表达系统在昆虫细胞中表达了完整的中国河北株丙型肝炎病毒（ＨＣＶ）结构蛋白。免疫印迹实验结果显示，表达产物中有一系列分子量不同、可以与ＨＣＶ抗体阳性病人血清反应的蛋白，表明结构蛋白被宿主细胞蛋白酶切割与加工，相应分别为２０ｋＤ的核心蛋白、３２ｋＤ糖基化的Ｅ１蛋白、４０ｋＤ的未糖化的Ｅ２蛋白和７０ｋＤ糖基化的Ｅ２蛋白，另有８０ｋＤ及１００ｋＤ的两组前体蛋白。利用表达产物检测慢性ＨＣＶ感染者血清，发现Ｅ２抗体检出率很高，而其中一部分核心抗体为阴性。A recombinant baculovirus containing the cDNA of the entire HCV structural region (derived from Chinese strain) was constructed and transfected into Sf9 insect cells by using Bac-to-Bac expression system. The expressed polyprotein was identified by HCV antibody positive human sera using Western blot and cleaved into a series of products with different sizes. 20kD, 32kD, 70kD, 40kD proteins were estimated as HCV Core, glycosylated E1, E2 and unglycoslated E2 proteins respectively. Two higher molecular weight proteins (80kD and 100kD) were also observed as some kinds of precursor of the structural polyprotein. HCV E2 antibody was more prevalent in chronic HCV infected human sera we examined. The antibodies to the expressed products (Core, E1) could be elicited in mice and the level was 1∶1280 and 1∶320 respectively by ELISA.

关 键 词：丙型肝炎病毒 结构基因 蛋白表达 杆状病毒 

分 类 号：R373.21[医药卫生—病原生物学] Q78[医药卫生—基础医学]