Characterization of the mouse insulin receptor gene promoter.
Characterization of the mouse insulin receptor gene promoter.
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小鼠胰岛素受体基因启动子的表征。
DOI：
10.1073/pnas.86.24.9732
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发表时间：
1989
影响因子：
11.1
通讯作者：
Lane,MD
中科院分区：
文献类型：
--
作者：
Sibley,E;Kastelic,T;Kelly,TJ;Lane,MD
分离小鼠胰岛素前受体基因的5'侧翼区，并定位最小启动子的5'边界。包含该基因的大于30个内切酶的基因组克隆含有启动子和外显子1和2，其在α亚基的氨基酸7的密码子处被约20个内切酶内含子中断。测定了含有766个碱基对的5'侧翼区和整个第一外显子的1.3-腺苷酸酶片段的核苷酸序列。通过S1核酸酶分析，将两个主要转录起始位点定位到位于翻译起始密码子上游469和424个核苷酸的位点。从小鼠3 T3-L1脂肪细胞cDNA文库中分离的胰岛素前受体cDNA的5'端对应于3'端最主要的转录起始位点。将连接在细菌氯霉素乙酰转移酶报告基因上游的前受体基因5'侧翼区的5'缺失突变体转染到3 T3-L1前脂肪细胞中，并测定启动子活性。最小启动子的5'边界（其指导出乎意料的高水平的报告基因表达）映射到3'最主要转录起始位点上游22个碱基对的区域。
The 5' flanking region of the mouse insulin proreceptor gene was isolated, and the 5' boundary of the minimal promoter was mapped. Genomic clones encompassing greater than 30 kilobases of the gene contain the promoter and exons 1 and 2 interrupted by an approximately 20-kilobase intron at the codon for amino acid 7 of the alpha subunit. The nucleotide sequence of a 1.3-kilobase fragment containing 766 base pairs of the 5' flanking region and the entire first exon was determined. Two major transcription start sites were mapped by S1 nuclease analysis to sites located 469 and 424 nucleotides upstream from the initiation codon for translation. The 5' terminus of an insulin proreceptor cDNA, isolated from a mouse 3T3-L1 adipocyte cDNA library, corresponds to the 3'-most major start site of transcription. The 5' deletion mutants of the 5' flanking region of the proreceptor gene, linked upstream of the bacterial chloramphenicol acetyltransferase reporter gene, were transfected into 3T3-L1 preadipocytes and assayed for promoter activity. The 5' boundary of the minimal promoter, which directs unexpectedly high levels of reporter gene expression, maps to a region 22 base pairs upstream from the 3'-most major transcription start site.