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小鼠醛氧化酶基因:5'侧翼区的分子克隆、染色体定位及功能特性分析

The mouse aldehyde oxidase gene: molecular cloning, chromosomal mapping and functional characterization of the 5'-flanking region.

作者信息

Demontis S, Kurosaki M, Saccone S, Motta S, Garattini E, Terao M

机构信息

Laboratory of Molecular Biology, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy.

出版信息

Biochim Biophys Acta. 1999 Dec 23;1489(2-3):207-22. doi: 10.1016/s0167-4781(99)00174-8.

Abstract

In this article, we report on the chromosome mapping and molecular cloning of the genetic locus encoding the mouse molybdo-iron/sulfur-flavoprotein aldehyde oxidase. The aldehyde oxidase locus maps to mouse chromosome 1 band C1-C2, as determined by fluorescence in situ hybridization experiments conducted on metaphase chromosomes. The gene is approximately 83 kb long and consists of 35 exons. The exon/intron boundaries are perfectly conserved relative to the corresponding human homolog and almost completely conserved relative to the mouse xanthine oxidoreductase gene. This further supports the concept that the aldehyde oxidase and xanthine oxidoreductase loci evolved from the same ancestral precursor by a gene duplication event. The position of a major transcription start site was defined by primer extension and RNase mapping analysis. The 5'-flanking region of the mouse aldehyde oxidase gene contains a functional and orientation-dependent promoter as well as several putative binding sites for known cell-specific and general transcription factors. Deletion analysis of the 5'-flanking region defines an approximately 470 bp DNA stretch which is necessary and sufficient for the transcription of the mouse aldehyde oxidase gene.

摘要

在本文中,我们报道了编码小鼠钼铁/硫黄素蛋白醛氧化酶的基因座的染色体定位和分子克隆。通过对中期染色体进行荧光原位杂交实验确定,醛氧化酶基因座定位于小鼠第1号染色体C1 - C2带。该基因长度约为83 kb,由35个外显子组成。外显子/内含子边界相对于相应的人类同源物完全保守,相对于小鼠黄嘌呤氧化还原酶基因几乎完全保守。这进一步支持了醛氧化酶和黄嘌呤氧化还原酶基因座通过基因复制事件从同一祖先前体进化而来的概念。通过引物延伸和核糖核酸酶图谱分析确定了主要转录起始位点的位置。小鼠醛氧化酶基因的5'侧翼区域包含一个功能性的、依赖于方向的启动子以及几个已知细胞特异性和一般转录因子的假定结合位点。对5'侧翼区域的缺失分析确定了一个约470 bp的DNA片段,该片段对于小鼠醛氧化酶基因的转录是必需且充分的。

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