| 丙型肝炎病毒NS_5B在Hub-7细胞中的转染与表达 |
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| 引用本文： | 房静远,EdwinOuyang,CatheineH.Wu,GeorgeY.Wu,邱德凯,曾民德,萧树东. 丙型肝炎病毒NS_5B在Hub-7细胞中的转染与表达[J]. 中华肝脏病杂志, 2000, 0(1) |
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| 作者姓名： | 房静远 EdwinOuyang CatheineH.Wu GeorgeY.Wu 邱德凯 曾民德 萧树东 |
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| 作者单位： | 上海第二医科大学附属仁济医院，上海市消化疾病研究所!200001，美国Connecticut大学医学院消化科，美国Connecticut大学医学院消化科，美国Connecticut大学医学院消化科，上海第二医科大学附属仁济医院，上海市消化疾病研究所!200001，上海第二医科大学附属仁济医院，上海市消化疾病研 |
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| 摘 要： | 目的建立丙型肝炎病毒（ＨＣＶ）ＮＳ５Ｂ表达的人肝细胞系。方法以脂质体共转染ＮＳ５ＳＢ基因人Ｈｕｂ－７细胞，以ＰＣＲ和Ｓｏｕｔｈｅｒｎ ｂｌｏｔ在ＤＮＡ水平检测转染的结果，以ｗｅｓｔｅｒｎ ｂｌｏｔ证实了Ｈｕｂ－７细胞中有ＮＳｕＢ的蛋白表达。结累在转染过ＮＳＳＢ质粒的细胞系中有ＮＳＳＢ基因的存在和ＨＣＶ－ＲＮＡ多聚酶的表达。结论我们在Ｈｕｂ－７细胞中建立了ＨＣＶ－ＲＮＡ多聚酶表达系统，它有助于研究ＨＣＶ复制机制和为基因治疗丙型肝炎打下基础。
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| 关 键 词： | 肝炎病毒 丙型 聚合酶链反应 转染 Huh-7细胞 |
Transfection and expression of HCV-NS_5B gene in Huh-7 cells |
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| FANG Jingyuan, Edwin Ouyang, Catheine H. Wu, et al. Transfection and expression of HCV-NS_5B gene in Huh-7 cells[J]. Chinese journal of hepatology, 2000, 0(1) |
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| Authors: | FANG Jingyuan Edwin Ouyang Catheine H. Wu et al |
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| Affiliation: | FANG Jingyuan, Edwin Ouyang, Catheine H. Wu, et al Shanghai institute of Digestive Disease,Shanghai Second Medical University,Shanghai 200001,China |
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| Abstract: | Objective Hepatitis C virus(HCV) is the major cause of blood-borne non-A, non-B hepatitis. An RNA-dependent RNA polymerase is encoded by HCV non-strUctural protein SB(NSSB) gene. A full-length HCV RNA has been transfected into human hepatoma cells, and HCV NS5B has been expressed in E.Coli purified. But human hepatocyte line with high expression of NSSB has not been established yet. Methods The NSSB gene has been transfected into Hub-7 cells by Lipofectamine. The results of transfection were confirmed by PCR and Southern blot analysis, and the level of the NSsB protein in Hub-7 cells was detected by Western blot analysis. Results There were the NSSB sequence and the expression of HCV-RNA polymerase in Hub-7 cells transfected NSSB plasmids. Conclusions We have established a HCV RNA polymerase expression system in Hub-7 cells that can be used to study the mechanism of HCV replication and to test gene therapy of hepatitis C. |
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| Keywords: | Hepatitis C virus Polymerase chain reachon Transfection Hub-7 cells |
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