Isolation of mouse THP gene promoter and demonstration of its kidney-specific activity in transgenic mice.
Isolation of mouse THP gene promoter and demonstration of its kidney-specific activity in transgenic mice.
复制标题
小鼠 THP 基因启动子的分离及其在转基因小鼠中的肾脏特异性活性的证明。
DOI：
10.1152/ajprenal.00297.2001
复制
发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Wu,Xue-Ru
中科院分区：
文献类型：
--
作者：
Zhu,Xinhua;Cheng,Jin;Gao,Jing;Lepor,Herbert;Zhang,Zhong-Ting;Pak,Joanne;Wu,Xue-Ru
Tamm-Horsfall蛋白（THP）是由肾脏上皮细胞合成的最丰富的尿蛋白，被认为在泌尿系统中发挥重要和多样的作用，包括肾水平衡、免疫抑制、尿结石形成和抑制细菌粘附。在本研究中，我们描述了小鼠THP基因的9.3kb，5′-区域的分离，并显示其近端589-bp，5′-侧翼序列与大鼠，牛和人类的高度保守性。我们还证明了使用转基因小鼠的方法，一个3.0 kb，近端5′-侧翼序列足以驱动肾脏特异性表达的异源报告基因。在肾脏内，转基因表达局限于内源性表达THP蛋白的肾小管，这表明在Henle袢和早期远曲小管的粗升支中具有特异性转基因活性。我们的研究结果建立了小鼠THP基因的肾脏和肾单位片段特异性表达。小鼠THP基因启动子在体内发挥作用，这将有助于进一步研究肾脏特异性基因调控的分子机制，并通过肾单位特异性基因靶向为更好地了解肾脏生理学和疾病提供新的分子工具。
Tamm-Horsfall protein (THP), the most abundant urinary protein synthesized by the kidney epithelial cells, is believed to play important and diverse roles in the urinary system, including renal water balance, immunosuppression, urinary stone formation, and inhibition of bacterial adhesion. In the present study, we describe the isolation of a 9.3-kb, 5′-region of the mouse THP gene and show the highly conserved nature of its proximal 589-bp, 5′-flanking sequence with that in rats, cattle, and humans. We also demonstrate using the transgenic mouse approach that a 3.0-kb, proximal 5′-flanking sequence is sufficient to drive the kidney-specific expression of a heterologous reporter gene. Within the kidney, transgene expression was confined to the renal tubules that endogenously expressed the THP protein, which suggests specific transgene activity in the thick ascending limb of the loop of Henle and early distal convoluted tubules. Our results establish the kidney- and nephron-segment-specific expression of the mouse THP gene. The availability of the mouse THP gene promoter that functions in vivo should facilitate additional studies of the molecular mechanisms of kidney-specific gene regulation and should provide new molecular tools for better understanding renal physiology and disease through nephron-specific gene targeting.