Cloning and characterization of the mouse glucokinase gene locus and identification of distal liver-specific DNase I hypersensitive sites.
Cloning and characterization of the mouse glucokinase gene locus and identification of distal liver-specific DNase I hypersensitive sites.
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DOI：
10.1006/geno.1995.9943
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发表时间：
1995-10
期刊：
影响因子：
4.4
通讯作者：
Catherine Postic;K. D. Niswender;J. Decaux;Ramine Parsa;K. Shelton;Betty Gouhot;C. C. Pettepher-C.
中科院分区：
文献类型：
--
作者：
Catherine Postic;K. D. Niswender;J. Decaux;Ramine Parsa;K. Shelton;Betty Gouhot;C. C. Pettepher-C.
我们克隆了一个83 kb的小鼠基因组DNA片段，其中包含完整的葡萄糖激酶（GK）基因。该基因的11个外显子的总距离为49 kb，外显子1 β和1 L相隔35 kb。确定了总共25，266 bp的DNA序列信息：相对于肝细胞转录起始位点，从约-9.2至约+15 kb（24，195 bp），相对于β细胞中的转录起始位点，从-335至+736 bp（1071 bp）。这些序列显示小鼠GK与大鼠和人GK的同一性> 94%。小鼠肝脏GK mRNA受禁食和再喂养的调节，大鼠也是如此。小鼠、大鼠和人类基因的上游和下游启动子区域的比对揭示了几个可能有助于转录调控的进化保守区域。然而，在转基因小鼠中的融合基因研究表明，肝细胞中转录起始位点附近的保守区域本身不足以在肝脏中进行位置无关性表达。使用DNase I对小鼠基因的48 kb区域的染色质结构进行分析，发现8个肝脏特异性超敏位点，其位置范围为肝脏转录起始位点上游0.1至36 kb。一个单一的，连续的DNA片段含有整个小鼠GK基因的可用性应允许GK的细胞特异性表达进行进一步的研究。
We cloned and characterized an 83-kb fragment of mouse genomic DNA containing the entire glucokinase (GK) gene. The 11 exons of the gene span a total distance of 49 kb, with exons 1 beta and 1L being separated by 35 kb. A total of 25,266 bp of DNA sequence information was determined: from approximately -9.2 to approximately +15 kb (24,195 bp), relative to the hepatocyte transcription start site, and from -335 to +736 bp (1071 bp), relative to the transcription start site in beta cells. These sequences revealed that mouse GK is > 94% identical to rat and human GK. Mouse hepatic GK mRNA is regulated by fasting and refeeding, as also occurs in the rat. Alignment of the upstream and downstream promoter regions of the mouse, rat, and human genes revealed several evolutionarily conserved regions that may contribute to transcriptional regulation. However, fusion gene studies in transgenic mice indicate that the conserved regions near the transcription start site in hepatocytes are themselves not sufficient for position-independent expression in liver. Analysis of the chromatin structure of a 48-kb region of the mouse gene using DNase I revealed eight liver-specific hypersensitive sites whose locations ranged from 0.1 to 36 kb upstream of the liver transcription start site. The availability of a single, contiguous DNA fragment containing the entire mouse GK gene should allow further studies of cell-specific expression of GK to be performed.