Cloning，Sequence Analysis and Expression of Partial HCV Nonstructural Protein 2，3 Gene

FAN Chao; GAO Jian-En; DAO Ji-Min

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August 2000

FAN Chao, GAO Jian-En and DAO Ji-Min. Cloning，Sequence Analysis and Expression of Partial HCV Nonstructural Protein 2，3 Gene[J]. Virologica Sinica, 2000, 15(4): 400-404.

Citation: FAN Chao, GAO Jian-En, DAO Ji-Min. Cloning，Sequence Analysis and Expression of Partial HCV Nonstructural Protein 2，3 Gene .VIROLOGICA SINICA, 2000, 15(4) : 400-404.

丙型肝炎病毒中国株非结构蛋白2～3部分基因cDNA克隆序列分析及表达

1.
北京医科大学人民医院肝病研究所，北京100044

摘要

丙型肝炎病毒 (ＨｅｐａｔｉｔｉｓＣｖｉｒｕｓ,ＨＣＶ)是输血后和许多社区获得性非甲非乙肝炎的主要致病因子。ＨＣＶ为单股正链ＲＮＡ病毒 ,其基因组长约 9.5Ｋｂ ,编码区含一个大开放读码框架 ,编码 30 1 0 30 33氨基酸残基的多蛋白前体 ,经宿主蛋白酶和病毒蛋白酶加...
- 丙型肝炎病毒
- , 非结构蛋白
- , 序列测定
- , 基因表达

Cloning，Sequence Analysis and Expression of Partial HCV Nonstructural Protein 2，3 Gene

1.
Institute of Hepatology, People'

Abstract

Complementary DNA of partial HCV NS2 NS3 protein gene which encodes viral Zn 2+ dependent metalloprotease and serine protease was amplified by RT nested PCR from serum of a Chinese patient with chronic hepatitis C.The product was digested with Eco R Ⅰ/ Xba Ⅰ and cloned into pcDNA3.The nucleotide sequence was determined.Comparison of NS2 NS3 with the reported typical isolates HCV 1,HCV J,HC C2,HCV J6,HCV J8 showed as 73.72%,90.20%,91.02%,64.56%,63.37% identity in nucleotide sequence and 83.24%,92.09%,93.13%, 70.88% ,67.86% identity in amino acid sequence respectively, so the isolate could be classified as HCV genotype Ⅱ. This gene fragment was cloned into pBV220 to construct recombinant expression vector pBV220 NS2 3.It was expressed efficiently in E.coli . The cloning and expression of HCV NS2 NS3 may contribute to the investigation of the relation between structure and function of viral encoded protease.
- Hepatitis C virus（HCV）
- , Nonstructural protein
- , Sequencing
- , Gene expressio

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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

Cloning，Sequence Analysis and Expression of Partial HCV Nonstructural Protein 2，3 Gene

1. Institute of Hepatology, People'

Abstract: Complementary DNA of partial HCV NS2 NS3 protein gene which encodes viral Zn 2+ dependent metalloprotease and serine protease was amplified by RT nested PCR from serum of a Chinese patient with chronic hepatitis C.The product was digested with Eco R Ⅰ/ Xba Ⅰ and cloned into pcDNA3.The nucleotide sequence was determined.Comparison of NS2 NS3 with the reported typical isolates HCV 1,HCV J,HC C2,HCV J6,HCV J8 showed as 73.72%,90.20%,91.02%,64.56%,63.37% identity in nucleotide sequence and 83.24%,92.09%,93.13%, 70.88% ,67.86% identity in amino acid sequence respectively, so the isolate could be classified as HCV genotype Ⅱ. This gene fragment was cloned into pBV220 to construct recombinant expression vector pBV220 NS2 3.It was expressed efficiently in E.coli . The cloning and expression of HCV NS2 NS3 may contribute to the investigation of the relation between structure and function of viral encoded protease.