| 中草药中胰高血糖素受体拮抗剂的高通量细胞筛选 |
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| 引用本文： | 万娟,许治良.中草药中胰高血糖素受体拮抗剂的高通量细胞筛选[J].华东师范大学学报(自然科学版),2007,2007(4):112-118. |
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| 作者姓名： | 万娟 许治良 |
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| 作者单位： | 华东师范大学,生命科学学院,上海市脑功能基因组学重点实验室,上海,200062 |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 摘 要： | 为筛选胰高血糖素受体拮抗剂，建立了一种高通量筛选胰高血糖素受体拮抗剂的细胞模型.利用PCR方法扩增得到人的胰高血糖素受体 cDNA，并连接到哺乳动物细胞表达载体pCDNA3.1上，构成质粒h glucagons-R/pCDNA3.1，将这种质粒与报告基因质粒(3×CRE/3×MRE/SRE-LUC)共转染到HEK293细胞，通过G418筛选，建立稳定的人胰高血糖素受体拮抗剂筛选细胞株.利用胰高血糖素受体内源激动剂探索和优化了筛选条件，筛选方法合理， Z′因子大于0.6，系统稳定，符合胰高血糖素受体拮抗剂的高通量筛选的要求.
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| 关 键 词： | 胰高血糖素受体 报告基因 拮抗剂 高通量药物筛选 胰高血糖素受体 报告基因 拮抗剂 高通量药物筛选 |
| 文章编号： | 1000-5641（2007）04-0112-07 |
| 收稿时间： | 2006-3-1 |
| 修稿时间： | 2006-03 |
High Throughput Screening Method of Identifying Antagonist for Glucagaon Receptor in Chinese Herbs(Chinese) |
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| WAN Juan,XU Zhi-liang.High Throughput Screening Method of Identifying Antagonist for Glucagaon Receptor in Chinese Herbs(Chinese)[J].Journal of East China Normal University(Natural Science),2007,2007(4):112-118. |
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| Authors: | WAN Juan XU Zhi-liang |
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| Institution: | Key Laboratory of Brain Functional Genomics of Shanghai, School of Life Science, East China Normal University, Shanghai 200062, China |
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| Abstract: | This paper generated a cell-based functional assay for human glucagon receptor antagonist screening. The human glucagon receptor cDNA was cloned into a mammalian expression vector pCDNA3.1 (h Glucagon receptor /pCDNA3.1). This plasmid was co-transfected with a reporter gene construct (3×CRE/3×MRE/SRE-LUC) in HEK293 cells. Stable cell line expressing both human glucagon receptor and the reporter gene was selected for compounds screening. The assay condition was optimized. Our results demonstrated that this cell-based assay can be optimized for high throughput screening and applied to identify antagonist for glucagon receptor in Chinese Herbs. |
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| Keywords: | glucagon receptor reporter gene antagonist high throughput screening |
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