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1.发明公开METHOD FOR REPRODUCING IN VITRO THE RNA-DEPENDENT RNA POLYMERASE AND TERMINAL NUCLEOTIDYL TRANSFERASE ACTIVITIES ENCODED BY HEPATITIS C VIRUS (HCV) 无效METHOD FOR RNA依赖性RNA聚合酶和丙型肝炎病毒末端核苷酸基转移酶活性的体外繁殖（HCV）
公开(公告)号：EP0842276A1
公开(公告)日：1998-05-20
申请号：EP96916289.0
申请日：1996-05-24
CPC分类号： C12N9/1241 , C12N9/127 , C12N2799/026
摘要： This is a method for reproducing in vitro the RNA-dependent RNA polymerase activity associated with hepatitis C virus. The method is characterized in that sequences contained in NS5B are used in the reaction mixture. The terminal nucleotidyl transferase activity, a further property of the NS5B protein, can also be reproduced using this method. The method takes advantage of the fact that the NS5B protein, either purified to apparent homogeneity or present in extracts of overproducing organisms, can catalyse the addition of ribonucleotides to the 3'-termini of exogenous or endogenous RNA molecules. The invention also relates to a composition of matter that comprises sequences contained in NS5B, and to the use of these compositions for the set up of an enzymatic test capable of selecting, for therapeutic purposes, compounds that inhibit the enzymatic activity associated with NS5B. The figure shows plasmids used in the method to produce hepatitis C virus RNA-dependent RNA polymerase and terminal nucleotidyl transferase in cultivated eukaryotic and prokaryotic cells.
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2.发明授权METHOD FOR REPRODUCING IN VITRO THE RNA-DEPENDENT RNA POLYMERASE AND TERMINAL NUCLEOTIDYL TRANSFERASE ACTIVITIES ENCODED BY HEPATITIS C VIRUS (HCV) 无效METHOD FOR RNA依赖性RNA聚合酶和丙型肝炎病毒末端核苷酸基转移酶活性的体外繁殖（HCV）
公开(公告)号：EP0842276B1
公开(公告)日：2004-01-21
申请号：EP96916289.0
申请日：1996-05-24
IPC分类号： C12N15/54 , C12N9/12 , C12Q1/48 , G01N33/573
CPC分类号： C12N9/1241 , C12N9/127 , C12N2799/026
摘要： This is a method for reproducing in vitro the RNA-dependent RNA polymerase activity associated with hepatitis C virus. The method is characterized in that sequences contained in NS5B are used in the reaction mixture. The terminal nucleotidyl transferase activity, a further property of the NS5B protein, can also be reproduced using this method. The method takes advantage of the fact that the NS5B protein, either purified to apparent homogeneity or present in extracts of overproducing organisms, can catalyse the addition of ribonucleotides to the 3'-termini of exogenous or endogenous RNA molecules. The invention also relates to a composition of matter that comprises sequences contained in NS5B, and to the use of these compositions for the set up of an enzymatic test capable of selecting, for therapeutic purposes, compounds that inhibit the enzymatic activity associated with NS5B. The figure shows plasmids used in the method to produce hepatitis C virus RNA-dependent RNA polymerase and terminal nucleotidyl transferase in cultivated eukaryotic and prokaryotic cells.
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