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Mouse connexin37: gene structure and promoter analysis

机译:小鼠连接蛋白37:基因结构和启动子分析

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Connexin37 (Cx37) is a subunit gap junction protein which exhibits limited expression in only a few cell types, predominantly in endothelial cells and in the lung. To begin to analyze Cx37 expression, we isolated a 1.6 kb mouse Cx37 cDNA from a mouse lung cDNA library and isolated corresponding mouse genomic clones from a bacterial artificial chromosome library. Sequencing and comparison of these clones showed that the Cx37 gene contained a short first exon, an 1.0 kb single intron and a second exon containing the complete coding region and 3'-untranslated region (UTR). The 5'-UTR of the mouse cDNA showed 70% identity to that of human Cx37. Primer extension experiments performed using mouse lung RNA gave two bands of sizes consistent with the transcription start site predicted from the cDNA. Sequence analysis showed that the regions flanking exon I contained a consensus 'TATA box' 43 bp 5' from the transcription start site preceded by several putative transcription factor binding sites and a 282 bp truncate L1Md interspersed element. Luciferase reporter gene transfections suggested that an area of 268 bp 5' from the first exon acted as a basal promoter for Cx37 and that there was a strong negative regulatory element in the intron.
机译:连接蛋白37(Cx37)是一种亚基间隙连接蛋白,仅在几种细胞类型中表达受限,主要在内皮细胞和肺中表达。为了开始分析Cx37的表达,我们从小鼠肺cDNA文库中分离了一个1.6 kb小鼠Cx37 cDNA,并从细菌人工染色体文库中分离了相应的小鼠基因组克隆。这些克隆的测序和比较表明,Cx37基因包含一个短的第一个外显子,一个1.0 kb的单个内含子和一个包含完整编码区和3'非翻译区(UTR)的第二个外显子。小鼠cDNA的5'-UTR与人Cx37的同源性为70%。使用小鼠肺RNA进行的引物延伸实验给出了两条带,其大小与从cDNA预测的转录起始位点一致。序列分析显示,外显子I侧翼区域从转录起始位点开始共有一个“ TATA框” 43 bp 5',之后是几个假定的转录因子结合位点和一个282 bp的截短的L1Md散布元件。萤光素酶报告基因的转染表明,第一个外显子的268 bp 5'区域充当Cx37的基础启动子,内含子中存在强的负调控元件。

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