水通道蛋白5（Aqp5）第一个内含子中的保守元件作为转录增强子发挥作用。
Conserved elements within first intron of aquaporin-5 (Aqp5) function as transcriptional enhancers.
作者信息
Flodby Per, Zhou Beiyun, Ann David K, Kim Kwang-Jin, Minoo Parviz, Crandall Edward D, Borok Zea
机构信息
Will Rogers Institute Pulmonary Research Center, Division of Pulmonary and Critical Care Medicine, Department of Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA.
出版信息
Biochem Biophys Res Commun. 2007 Apr 27;356(1):26-31. doi: 10.1016/j.bbrc.2007.02.076. Epub 2007 Feb 23.
A 4.3 kb rat aquaporin-5 (Aqp5) promoter that directs lung and salivary cell-specific expression in vitro directs low level expression of a GFP reporter in lungs of transgenic mice. Alignment of rat, mouse, and human AQP5 genomic sequences identified a highly conserved region in the 3' portion of intron 1, here termed ci1. To investigate the role of ci1 in Aqp5 expression, transient transfections were undertaken in AQP5-expressing mouse lung epithelial (MLE-15) and rat salivary (Pa-4) cells and AQP5-non-expressing NIH/3T3 cells. A 536 bp ci1 fragment enhanced transcriptional activity of the rat Aqp5 minimal promoter specifically in MLE-15 cells in an orientation-independent manner. Enhancer activity was Aqp5 promoter-specific, since no increase in activity was detected with the TK promoter. These results suggest that expression of transgenes in mouse lungs under direction of the 4.3 kb rat Aqp5 promoter may be augmented by inclusion of ci1 in transgenic constructs.
一个4.3 kb的大鼠水通道蛋白5（Aqp5）启动子在体外可指导肺和唾液腺细胞特异性表达，该启动子在转基因小鼠的肺中指导绿色荧光蛋白（GFP）报告基因的低水平表达。大鼠、小鼠和人类AQP5基因组序列的比对在第1内含子的3'部分鉴定出一个高度保守的区域，在此称为ci1。为了研究ci1在Aqp5表达中的作用，在表达AQP5的小鼠肺上皮（MLE-15）细胞、大鼠唾液腺（Pa-4）细胞和不表达AQP5的NIH/3T3细胞中进行了瞬时转染。一个536 bp的ci1片段以方向独立的方式特异性增强了大鼠Aqp5最小启动子在MLE-15细胞中的转录活性。增强子活性是Aqp5启动子特异性的，因为用胸苷激酶（TK）启动子未检测到活性增加。这些结果表明，在转基因构建体中包含ci1可能会增强4.3 kb大鼠Aqp5启动子指导下的转基因在小鼠肺中的表达。
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