文献类型：期刊文献

中文题名：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

英文题名：Optimization of Androgen-Regulatable,Kidney Proximal Renal Tubule Epithelial Cell Specific Targeted Kap147/HAGT-Kpn Hybrid Promoter

作者：孟军[1];刘飞[1];范立强[1];朱庆庆[1];吴宝杰[1];左丽君[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

年份：2014

卷号：40

期号：1

起止页码：47

中文期刊名：华东理工大学学报（自然科学版）

外文期刊名：Journal of East China University of Science and Technology

收录：CSTPCD;;Scopus;北大核心:【北大核心2011】；CSCD:【CSCD2013_2014】；

基金：国家自然科学基金(81071252)

语种：中文

中文关键词：肾脏;雄激素调节蛋白;杂合启动子;雄激素应答元件

外文关键词：kidney; androgen-regulated protein; hybrid promoter; androgen response element

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20nmol／LDHT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20nmol／LDHT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。
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.