A nonisotopic assay method for hepatitis C virus NS5B polymerase |
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| Authors: | Park Chanhee Kee Younghoon Park Jungchan Myung Heejoon |
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| Affiliation: | Department of Bioscience and Biotechnology, Hankuk University of Foreign Studies, Kyung-Gi Do, South Korea. |
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| 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. |
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