A nonisotopic assay method for hepatitis C virus NS5B polymerase

被引：11

作者：

Park, C

Kee, Y

Park, J

Myung, H ^{[1
]}

机构：

[1] Hankuk Univ Foreign Studies, Dept Biosci & Biotechnol, Kyonggi Do, South Korea

[2] ChemBioNex Co Ltd, Yongin 449850, Kyung Gi Do, South Korea

来源：

JOURNAL OF VIROLOGICAL METHODS | 2002年 / 101卷 / 1-2期

关键词：

hepatitis C virus; NS5B polymerase; covalink module; nonisotopic assay;

D O I：

10.1016/S0166-0934(01)00443-8

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The RNA-dependent RNA polymerase of hepatitis C virus (HCV) is responsible for replication of genomic RNA. A novel nonisotopic assay method is described for detecting its enzymatic activity. The 5' end of the in vitro-transcribed template RNA was attached covalently to the surface of a Covalink module using carbodiimide condensation. The RNA strand containing the 3' untranslated region (3' UTR) of HCV at its 3' end was free in the solution. A purified NS5B polymerase and NTPs along with biotin-labeled UTP were added to this module and the polymerization activity could be detected colorimetrically with streptavidin-conjugated alkaline phosphatase. (C) 2002 Elsevier Science B.V. All rights reserved.

引用

页码：211 / 214

页数：4

共 20 条

[11]

Lohmann V, 2000, J VIRAL HEPATITIS, V7, P167

[12] Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line [J].