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J Virol Methods. 2002 Mar;101(1-2):211-4.

A nonisotopic assay method for hepatitis C virus NS5B polymerase.

Author information

1
Department of Bioscience and Biotechnology, Hankuk University of Foreign Studies, Kyung-Gi Do, South Korea.

Abstract

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.

PMID:
11849700
[Indexed for MEDLINE]

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