| 丙型肝炎病毒非结构蛋白NS5A反式激活基因NS5A-TP9启动子序列的确定及转录活性的鉴定 |
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| 引用本文： | 巨立中,钟彦伟,成军,王建军,洪源. 丙型肝炎病毒非结构蛋白NS5A反式激活基因NS5A-TP9启动子序列的确定及转录活性的鉴定[J]. 中西医结合肝病杂志, 2003, 13(6): 357-359 |
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| 作者姓名： | 巨立中 钟彦伟 成军 王建军 洪源 |
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| 作者单位： | 1. 中国人民解放军第273医院,新疆,库尔勒,841000
2. 中国人民解放军第302医院传染病研究所基因治疗研究中心 |
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| 基金项目： | 国家自然科学基金(No.C39970674,No.C03011402,No.C39900130,No.C30070689) |
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| 摘 要： | 目的：阐明丙型肝炎病毒(HCV)非结构蛋白NS5A(NS5A)反式激活基因NS5A-TP9表达的调控机制。方法：选取NS5A-TP9基因翻译起始密码子ATG上游661bp的基因组DNA序列作为启动子序列，应用聚合酶链反应(PCR)技术，以肝母细胞瘤细胞系HepG2基因组DNA为模板，扩增该启动子DNA片段，将其克隆至pCAT3中，构建pCAT3-NS5A-TP9-promoter报告基因表达载体，以该质粒转染HepG2细胞，用酶联免疫吸附法(ELISA)检测报告基因编码产物氯霉素乙酰转移酶(CAT)的表达活性。结果：发现质粒pCAT3-NS5A-TP9-promoter具有指导报告基因CAT转录表达的启动子活性，CAT的吸光度(A值)是缺乏启动子序列对照质粒pCAT3的13．9倍。结论：本研究克隆的启动子DNA序列具有指导下游基因转录的启动子活性，这一结果为研究新基因NS5A-TP9转录表达的调节机制，进一步阐明丙型肝炎病毒非结构蛋白NS5A的作用机制奠定了基础。
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| 关 键 词： | 丙型肝炎病毒 非结构蛋白 反式激活基因 启动子 转录活性 聚合酶链反应 克隆 |
| 修稿时间： | 2003-08-25 |
Identification and Evaluation of Promoter Sequence and Its Transcription Activation of the Human Gene 9 Transactivated by Nonstructural Protein 5A of Hepatitis C Virus |
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| JU Lizhong,ZHONG Yanwei,CHENG Jim,et al.. Identification and Evaluation of Promoter Sequence and Its Transcription Activation of the Human Gene 9 Transactivated by Nonstructural Protein 5A of Hepatitis C Virus[J]. Chinese Journal of Integrated Traditonal and Western Medicine on Liver Diseases, 2003, 13(6): 357-359 |
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| Authors: | JU Lizhong ZHONG Yanwei CHENG Jim et al. |
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| Affiliation: | JU Lizhong,ZHONG Yanwei,CHENG Jim,et al. 1. The 273 Hospital of Chinese PLA |
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| Abstract: | Objective: To clarify the expression and regulation mechanism of the new target gene (NS5A-TP9) transacti-vated by nonstructural protein 5A (NS5A) of hepatitis C virus (HCV) .Methods: Six hundred and sixty-one base pairs (bp) up-stream of the translation start site was selected as promoter sequence which was amplified from hepatoblastoma cell line-HepG 2 cell genomic DNA by polymerase chain reaction (PCR) .The amplified product was cloned into pCAT3 vector.The HepG2 cell line was tranfected by pCAT3-NS5ATP9-promoter.The CAT activity was detected by an enzyme-linked immunosorbent assay (ELISA) kit. Results: We found pCAT3-NS5ATP9-promoter had high activity of CAT expression.The expression of CAT was 13.9 times as higher as the negative control plasmid pCAT3-basic.Conclusion: These results indicated that the pCAT3-NS5ATP9-promoter has promoter activity. |
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| Keywords: | Molecular Biology Hepatitis C Virus Transactivation Gene Cloning Promoter |
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