A nonisotopic assay method for hepatitis C virus NS5B polymerase

Chanhee Park; Younghoon Kee; Jungchan Park; Heejoon Myung

doi:10.1016/S0166-0934(01)00443-8

A nonisotopic assay method for hepatitis C virus NS5B polymerase

Chanhee Park
, Younghoon Kee
, Jungchan Park
, Heejoon Myung

Department of New Biology

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

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.

Original language	English
Pages (from-to)	211-214
Number of pages	4
Journal	Journal of Virological Methods
Volume	101
Issue number	1-2
DOIs	https://doi.org/10.1016/S0166-0934(01)00443-8
State	Published - 2002

Keywords

Covalink module
Hepatitis C virus
NS5B polymerase
Nonisotopic assay

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/S0166-0934(01)00443-8

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title = "A nonisotopic assay method for hepatitis C virus NS5B polymerase",

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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T1 - A nonisotopic assay method for hepatitis C virus NS5B polymerase

AU - Park, Chanhee

AU - Kee, Younghoon

AU - Park, Jungchan

AU - Myung, Heejoon

PY - 2002

Y1 - 2002

N2 - 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.

AB - 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.

KW - Covalink module

KW - Hepatitis C virus

KW - NS5B polymerase

KW - Nonisotopic assay

UR - https://www.scopus.com/pages/publications/0036175506

U2 - 10.1016/S0166-0934(01)00443-8

DO - 10.1016/S0166-0934(01)00443-8

M3 - Article

C2 - 11849700

AN - SCOPUS:0036175506

SN - 0166-0934

VL - 101

SP - 211

EP - 214

JO - Journal of Virological Methods

JF - Journal of Virological Methods

IS - 1-2

ER -

A nonisotopic assay method for hepatitis C virus NS5B polymerase

Abstract

Keywords

UN SDGs

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