| 人酰基辅酶A:胆固醇酰基转移酶1基因P7启动子调控的报告基因载体的构建及鉴定 |
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| 引用本文： | 葛晶,成蓓,何平,文晖,陆菡,陈心,曾永利. 人酰基辅酶A:胆固醇酰基转移酶1基因P7启动子调控的报告基因载体的构建及鉴定[J]. 生物医学工程学杂志, 2008, 25(6) |
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| 作者姓名： | 葛晶 成蓓 何平 文晖 陆菡 陈心 曾永利 |
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| 作者单位： | 华中科技大学,同济医学院,附属协和医院,老年病科,武汉,430022 |
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| 基金项目： | 国家自然科学基金资助项目 |
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| 摘 要： | 应用PCR方法从人单核细胞系THP-1基因组DNA扩增人酰基辅酶A:胆固醇酰基转移酶1(acyl coenzyme A: cholesterol acyltransferase1, ACAT1)基因 P7启动子全长片段,进行TA克隆,经双酶切、测序鉴定阳性克隆.将该基因亚克隆至荧光素酶报告基因载体pGL3-Enhancer,用DEAE-葡聚糖(DEAE-dextran)法将重组质粒pGL3E-P7瞬时转染入单核细胞THP-1,检测其活性,证明pGL3E-P7能在体外表达荧光素酶.人ACAT1基因P7启动子调控的荧光素酶报告基因表达载体的成功构建,为研究动脉粥样硬化过程中ACAT1基因的转录调控机制提供新手段.
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| 关 键 词： | 动脉粥样硬化 酰基辅酶A 胆固醇酰基转移酶1 启动子 荧光素酶 |
Construction and Identification of Recombinant Firefly Luciferase Report Vector Containing Human Acyl Coenzyme A: Cholesterol Acyltransferase 1 Gene P7 Promoter |
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| Ge Jing,Cheng Bei,He Ping,Wen Hui,Lu Han,Chen Xin,Zeng Yongli. Construction and Identification of Recombinant Firefly Luciferase Report Vector Containing Human Acyl Coenzyme A: Cholesterol Acyltransferase 1 Gene P7 Promoter[J]. Journal of biomedical engineering, 2008, 25(6) |
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| Authors: | Ge Jing Cheng Bei He Ping Wen Hui Lu Han Chen Xin Zeng Yongli |
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| Affiliation: | Department of Gerontology;Union Hospital;Tongji Medical College of Huazhong University of Science and Technology;Wuhan 430022;China |
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| Abstract: | The DNA segment of the human acyl coenzyme A: cholesterol acyltransferase1(ACAT1) gene P7 promoter was amplified by PCR from human monocytic leukemia cell line(THP-1) and cloned to TA vector,then the positive clone was confirmed by restriction enzymes and sequencing.The targeted segment was subcloned to Firefly luciferase report vector pGL3-Enhancer.The recombinant plasmid pGL3EP7 was transfected transiently into THP-1,then the expression of luciferase could be detected in THP-1 by pGL3E-P7 transfection.We ... |
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