| 乙型肝炎病毒基因组中前-前-S-编码基因启动子序列的确定及转录活性的鉴定 |
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| 引用本文： | 杨倩,董菁,成军,刘妍,洪源,王建军,张树林. 乙型肝炎病毒基因组中前-前-S-编码基因启动子序列的确定及转录活性的鉴定[J]. 解放军医学杂志, 2003, 28(9): 761-762,765 |
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| 作者姓名： | 杨倩 董菁 成军 刘妍 洪源 王建军 张树林 |
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| 作者单位： | 100039,北京,解放军第302医院传染病研究所基因治疗研究中心;100039,北京,解放军第302医院传染病研究所基因治疗研究中心;100039,北京,解放军第302医院传染病研究所基因治疗研究中心;100039,北京,解放军第302医院传染病研究所基因治疗研究中心;100039,北京,解放军第302医院传染病研究所基因治疗研究中心;100039,北京,解放军第302医院传染病研究所基因治疗研究中心;100039,北京,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 (编号C39970 674,C0 30 1 1 4 0 2 ,C30 0 70 689,C3990 0 1 30 )，军队“十五”科技攻关青年基金 (编号 0 1Q1 38)，军队“十五”科技攻关面上项目 (编号 0 1MB1 35)，军队留学归国人员启动基金 (编号 98H0 38)资助课题 |
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| 摘 要： | 为了解前-前-S编码区ORF上游是否具有启动子活性，选取其起始密码子ATG上游277bp的DNA序列，将其克隆至报告基因表达载体pCAT3中，构建pCAT3-前-前-S-promoter表达载体，以该质粒转染HepG2细胞，用酶联免疫吸附法(ELISA)检测氯霉素乙酰转移酶(CAT)的表达活性。结果发现，质粒pCAT3-前-前-S-pmrnoter能够激活CAT的表达，吸光度(A值)是pCAT3-basic的10倍，说明pCAT3-前-前-S-promoter具有启动子活性，为在HBV基因组DNA序列中界定、研究新的前-前-SORF提供了直接的实验依据。
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| 关 键 词： | 肝炎病毒 乙型 前-前-S基因 启动子 |
DEFINITION OF PRE-PRE-S PROMOTER SEQUENCE FROM HEPATITIS B VIRUS GENOME AND IDENTIFICATION OF ITS TRANSCRIPTION ACTIVITY |
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| Yang Qian,Dong Jing,Cheng Jun et al. Gene Therapy Research Center. DEFINITION OF PRE-PRE-S PROMOTER SEQUENCE FROM HEPATITIS B VIRUS GENOME AND IDENTIFICATION OF ITS TRANSCRIPTION ACTIVITY[J]. Medical Journal of Chinese People's Liberation Army, 2003, 28(9): 761-762,765 |
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| Authors: | Yang Qian Dong Jing Cheng Jun et al. Gene Therapy Research Center |
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| Affiliation: | Yang Qian,Dong Jing,Cheng Jun et al. Gene Therapy Research Center,Institute of Infectious Diseases,302 Hospital of PLA,Beijing 100039,China |
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| Abstract: | To investigate the promoter activity of pre-pre-S gene, which was found in five clones of hepatitis B virus (HBV) genome from 2 patients with chronic HBV infection, promoter DNA sequence was amplified from HBV-DNA by polymerase chain reaction (PCR).The amplified product was cloned into pCAT3 reporter vector. The HepG2 cells were transfected by pCAT3-pre-pre-S-p. The CAT activity was detected by an enzyme-linked immunosorbent assay (ELISA) kit. It was found that pCAT3-pre-pre-S-p had higher activity of CAT which was 10 fold over that of pCAT3-basic. This result implicated that pCAT3-pre-pre-S-p had promoter activity. |
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| Keywords: | hepatitis B virus pre-pre-S-region promoter |
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