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首页> 外文期刊>Molecular and Cellular Biology >Alu sequence involvement in transcriptional insulation of the keratin 18 gene in transgenic mice.
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Alu sequence involvement in transcriptional insulation of the keratin 18 gene in transgenic mice.

机译：Alu序列参与转基因小鼠中角蛋白18基因的转录绝缘。

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The human keratin 18 (K18) gene is expressed in a variety of adult simple epithelial tissues, including liver, intestine, lung, and kidney, but is not normally found in skin, muscle, heart, spleen, or most of the brain. Transgenic animals derived from the cloned K18 gene express the transgene in appropriate tissues at levels directly proportional to the copy number and independently of the sites of integration. We have investigated in transgenic mice the dependence of K18 gene expression on the distal 5' and 3' flanking sequences and upon the RNA polymerase III promoter of an Alu repetitive DNA transcription unit immediately upstream of the K18 promoter. Integration site-independent expression of tandemly duplicated K18 transgenes requires the presence of either an 825-bp fragment of the 5' flanking sequence or the 3.5-kb 3' flanking sequence. Mutation of the RNA polymerase III promoter of the Alu element within the 825-bp fragment abolishes copy number-dependent expression in kidney but does not abolish integration site-independent expression when assayed in the absence of the 3' flanking sequence of the K18 gene. The characteristics of integration site-independent expression and copy number-dependent expression are separable. In addition, the formation of the chromatin state of the K18 gene, which likely restricts the tissue-specific expression of this gene, is not dependent upon the distal flanking sequences of the 10-kb K18 gene but rather may depend on internal regulatory regions of the gene.
机译：人角蛋白18（K18）基因在多种成人的简单上皮组织中表达，包括肝，肠，肺和肾，但通常在皮肤，肌肉，心脏，脾脏或大部分大脑中不存在。源自克隆的K18基因的转基因动物在适当的组织中以与拷贝数成正比且与整合位点无关的水平表达转基因。我们已经在转基因小鼠中研究了K18基因表达对5'和3'末端侧翼序列以及紧靠K18启动子上游的Alu重复DNA转录单元的RNA聚合酶III启动子的依赖性。串联复制的K18转基因的整合位点非依赖表达需要存在5'侧翼序列的825-bp片段或3.5-kb 3'侧翼序列的存在。在不存在K18基因的3'侧翼序列的情况下进行测定时，825-bp片段中Alu元件的RNA聚合酶III启动子的突变消除了肾脏中拷贝数依赖性的表达，但没有消除整合位点依赖性的表达。整合位点无关表达和拷贝数依赖性表达的特征是可分离的。此外，K18基因染色质状态的形成可能会限制该基因的组织特异性表达，它并不依赖于10kb K18基因的远端侧翼序列，而可能依赖于K18基因的内部调控区。基因。

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