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    孟军, 刘飞, 范立强, 朱庆庆, 吴宝杰, 左丽君. 雄激素可调节的肾脏近端肾小管上皮细胞靶向杂合启动子的优化[J]. 华东理工大学学报(自然科学版), 2014, (1): 47-52.
    引用本文: 孟军, 刘飞, 范立强, 朱庆庆, 吴宝杰, 左丽君. 雄激素可调节的肾脏近端肾小管上皮细胞靶向杂合启动子的优化[J]. 华东理工大学学报(自然科学版), 2014, (1): 47-52.
    MENG Jun, LIU Fei, FAN Li-qiang, ZHU Qing-qing, WU Bao-jie, ZUO Li-jun. Optimization of Androgen Regulatable, Kidney Proximal Renal Tubule Epithelial Cell Specific Targeted Kap147/HAGT Kpn Hybrid Promoter[J]. Journal of East China University of Science and Technology, 2014, (1): 47-52.
    Citation: MENG Jun, LIU Fei, FAN Li-qiang, ZHU Qing-qing, WU Bao-jie, ZUO Li-jun. Optimization of Androgen Regulatable, Kidney Proximal Renal Tubule Epithelial Cell Specific Targeted Kap147/HAGT Kpn Hybrid Promoter[J]. Journal of East China University of Science and Technology, 2014, (1): 47-52.

    雄激素可调节的肾脏近端肾小管上皮细胞靶向杂合启动子的优化

    Optimization of Androgen Regulatable, Kidney Proximal Renal Tubule Epithelial Cell Specific Targeted Kap147/HAGT Kpn Hybrid Promoter

    • 摘要: 为研究肾脏雄激素调节蛋白杂合启动子Kap147/HAGT Kpn的细胞特异性,将HAGT Kpn片段克隆到pGL3 Kap147载体质粒上,通过细胞转染和荧光素酶活性测定,证实Kap147/HAGT Kpn杂合启动子只在肾小管上皮细胞来源的OK、LLC MK2细胞中具有活性,经20 nmol/L DHT处理后其活性增加了7~8倍。依据雄激素应答原件的保守性,将不同长度的HAGT Kpn亚片段克隆到pGL3 Kap147载体质粒上,经20 nmol/L DHT处理后,KpnI StuI和MscI KpnI两个片段能够使Kap147活性增强1~2倍,由此表明Kap147/HAGT Kpn杂合启动子依然具有组织细胞特异性且受雄激素调控,KpnI StuI和MscI KpnI两个片段承担了雄激素的调控作用。

       

      Abstract: Abstract: To study the cell specificity of the Kap147/HAGT Kpn hybrid promoter enhancer combination,the HAGT Kpn fragment was cloned into pGL3 Kap147 vector. Luciferase activity assay after cell transient transfection showed that Kap147/HAGT Kpn hybrid promoter only exhibited activity in OK and LLC ML2 cells that derived from epithelial cells of proximal convoluted tubules, and its activity increased by 7 to 8 fold treated with 20 nmol/L DHT. In order to optimize the hybrid promoter enhancer combination, truncated HAGT Kpn fragments were cloned into pGL3 Kap147 vector. Cell experiments showed that KpnI StuI and MscI KpnI fragments increased expression of Kap147 promoter 1 to 2 fold treated with 20 nmol/L DHT. Kap147/HAGT Kpn hybrid promoter is tissue and cell specific and androgen regulated, while KpnI StuI and MscI KpnI fragments are responsible for the androgen regulatory effect.

       

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