Expression and Localization of the HCV Structural Protein E1 in Insect Cells

MA Jia; WANG Rui-qing; SONG Jian-hua; LIANG Chang-yong; CHEN Xin

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October 2005

MA Jia, WANG Rui-qing, SONG Jian-hua, LIANG Chang-yong and CHEN Xin. Expression and Localization of the HCV Structural Protein E1 in Insect Cells[J]. Virologica Sinica, 2005, 20(5): 480-484.

Citation: MA Jia, WANG Rui-qing, SONG Jian-hua, LIANG Chang-yong, CHEN Xin. Expression and Localization of the HCV Structural Protein E1 in Insect Cells .VIROLOGICA SINICA, 2005, 20(5) : 480-484.

丙型肝炎病毒结构蛋白E1在昆虫细胞中的表达及定位

马佳 ^1,2 ,
王瑞青 ¹ ,
宋建华 ^1,2 ,
梁昌镛 ^1,2 ,
陈新文

1.
1.中国科学院武汉病毒研究所病毒学国家重点实验室湖北武汉 2.中国科学院研究生院北京

摘要

本文在大肠杆菌中表达了与GST融合无跨膜区的丙型肝炎病毒(Hepatitis C Virus,HCV)E1蛋白,并通过免疫兔制备了兔抗E1的抗血清。然后利用Bac-to-Bac杆状病毒表达系统构建了含有HCV结构蛋白E1基因的重组杆状病毒vAcHCVE1。通过Western blot分析,E1蛋白在Sf9细胞中表达分子量大小为30kDa大于预测的20kDa,表明存在翻译后修饰如糖基化等。通过Confocal显微镜观察当感染48h后E1蛋白定位在细胞质和细胞膜上。
- 丙型肝炎病毒
- , E1
- , 表达
- , Sf9细胞
- , 激光共聚焦显微镜

Expression and Localization of the HCV Structural Protein E1 in Insect Cells

Abstract

In this work,the truncated Hepatitis C virus(HCV) structural protein E1 fused with GST was expressed in E.coli.The fusion protein was used as an antigen to immunize rabbit to produce the antibody for detection.To express HCV E1 in insect cells,the recombinant baculovirus(vAcHCVE1) containing the HCV full length E1 gene was constructed by Bac-to-Bac recombinant baculovirus expression system.It was proved to be able to produce HCV structural protein E1 in insect cell Sf9.The expression protein was detected by Western blot analysis.The size of the product was 30 kDa,which was bigger than the predicted size of 20 kDa,implicating post-translational modification such as glycosylation.By using confocal microscope,it was observed that the E1 protein was localized in the cytoplasm and cellular membrane 48h after inflection.
- Hepatitis C virus（HCV）
- , E1
- , Expression
- , Sf9
- , Confocal microscope

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Expression and Localization of the HCV Structural Protein E1 in Insect Cells

Abstract: In this work,the truncated Hepatitis C virus(HCV) structural protein E1 fused with GST was expressed in E.coli.The fusion protein was used as an antigen to immunize rabbit to produce the antibody for detection.To express HCV E1 in insect cells,the recombinant baculovirus(vAcHCVE1) containing the HCV full length E1 gene was constructed by Bac-to-Bac recombinant baculovirus expression system.It was proved to be able to produce HCV structural protein E1 in insect cell Sf9.The expression protein was detected by Western blot analysis.The size of the product was 30 kDa,which was bigger than the predicted size of 20 kDa,implicating post-translational modification such as glycosylation.By using confocal microscope,it was observed that the E1 protein was localized in the cytoplasm and cellular membrane 48h after inflection.