| 黑色素皮质素受体激动剂的高通量筛选模型研究 |
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| 引用本文： | 郑旭煦,欧阳克清,高虹,许治良,胡应和,蔡绍皙. 黑色素皮质素受体激动剂的高通量筛选模型研究[J]. 生物化学与生物物理进展, 2004, 31(1): 36-40 |
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| 作者姓名： | 郑旭煦 欧阳克清 高虹 许治良 胡应和 蔡绍皙 |
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| 作者单位： | 1. 重庆大学生物工程学院,生物力学与组织工程教育部重点实验室,重庆,400044;重庆工商大学环境与生物工程学院,重庆,400033
2. 重庆大学生物工程学院,生物力学与组织工程教育部重点实验室,重庆,400044 |
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| 基金项目： | 重庆市科委攻关项目（渝科发计字［2002］1号）和重庆市计委重点高新技术项目（渝计委计［2001］1192号）. |
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| 摘 要： | 为了建立黑色素皮质素受体（MC4R）激动剂的高通量筛选方法，将人的MC4R基因质粒（hMC4R/pCDNA3.1）与报告基因质粒（3×CRE/3×MRE/SRE-LUC）按1∶5的比例共转染到HEK293细胞，通过G418筛选，建立了稳定的MC4R激动剂筛选细胞株.利用MC4R内源激动剂α-MSH探索和优化了每孔接种细胞数目、激动剂孵育时间、溶剂DMSO终浓度、荧光素酶底物浓度等筛选条件，建立了可靠的筛选方法.实验表明：当细胞数目为4×104个/孔,激动剂孵育时间为8 h,每孔DMSO终浓度小于1％和α-MSH终浓度为1 μmol/L时，系统Z'-因子接近0.7，能够用于MC4R激动剂的高通量筛选.
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| 关 键 词： | 黑色素皮质素受体(MC4R)，报告基因，激动剂，高通量筛选 |
| 收稿时间： | 2003-05-26 |
| 修稿时间： | 2003-07-28 |
High Throughput Screening Method of Identifying Agonist for Melanocortin 4 Receptor |
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| ZHENG Xu-Xu,OUYANG Ke-Qing,GAO Hong,XU Zhi-Liang,HU Ying-He and CAI Shao-Xi. High Throughput Screening Method of Identifying Agonist for Melanocortin 4 Receptor[J]. Progress In Biochemistry and Biophysics, 2004, 31(1): 36-40 |
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| Authors: | ZHENG Xu-Xu OUYANG Ke-Qing GAO Hong XU Zhi-Liang HU Ying-He CAI Shao-Xi |
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| Affiliation: | College of Bioengineering, Key Laboratory of Biomechanics & Tissue Engineering of Ministry of Education, Chongqing University, Chongqing 400044, China;College of Environment & Biological Engineering, Chongqing Technology & Business University, Chongqing 4;College of Bioengineering, Key Laboratory of Biomechanics & Tissue Engineering of Ministry of Education, Chongqing University, Chongqing 400044, China;College of Bioengineering, Key Laboratory of Biomechanics & Tissue Engineering of Ministry of Education, Chongqing University, Chongqing 400044, China;College of Bioengineering, Key Laboratory of Biomechanics & Tissue Engineering of Ministry of Education, Chongqing University, Chongqing 400044, China;College of Bioengineering, Key Laboratory of Biomechanics & Tissue Engineering of Ministry of Education, Chongqing University, Chongqing 400044, China;College of Bioengineering, Key Laboratory of Biomechanics & Tissue Engineering of Ministry of Education, Chongqing University, Chongqing 400044, China |
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| Abstract: | |
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| Keywords: | melanocortin 4 receptor reporter gene agonist high throughput screening (HTS) |
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