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中草药中胰高血糖素受体拮抗剂的高通量细胞筛选 被引量：4

High Throughput Screening Method of Identifying Antagonist for Glucagaon Receptor in Chinese Herbs
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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. 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.
作者 万娟 许治良
机构地区 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
出处 《华东师范大学学报（自然科学版）》 CAS CSCD 北大核心 2007年第4期112-118,共7页 Journal of East China Normal University(Natural Science)
基金 国家科技部863计划项目(2003AA221061)
关键词 胰高血糖素受体 报告基因 拮抗剂 高通量药物筛选 glucagon receptor reporter gene antagonist high throughput screening
分类号 R965 [医药卫生—药理学]
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参考文献15

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共引文献34

同被引文献66

引证文献4

二级引证文献6

华东师范大学学报（自然科学版）
2007年 第4期

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