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首页> 外文期刊>Molecular and Cellular Biology >Molecular cloning of the murine adenosine deaminase gene from a genetically enriched source: identification and characterization of the promoter region.
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Molecular cloning of the murine adenosine deaminase gene from a genetically enriched source: identification and characterization of the promoter region.

机译：从遗传丰富的来源克隆鼠腺苷脱氨酶基因：启动子区域的鉴定和表征。

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摘要

A genomic library was prepared with DNA from a genetically enriched mouse cell line in which amplified copies of the adenosine deaminase (ADA) gene account for over 5% of the genome. Overlapping cosmid clones encompassing the entire ADA structural gene were isolated from this genomic library and used for subsequent structural and functional analyses. Nuclease protection and primer extension analyses served to identify the location of multiple transcription initiation sites at the 5' end of the structural gene. Promoter activity was found by functional analyses to reside within a 240-base-pair fragment which contains the transcription initiation sites. Sequences upstream of the transcription initiation sites are very G + C rich (77%) and include a 22 nucleotide stretch of deoxyguanylate residues and two potential Sp1 transcription factor-binding sites. Comparison of the mouse and human ADA gene promoters revealed the presence of several regions that are highly conserved with regard to both sequence content and location and may represent genetic elements which are involved in ADA gene expression.
机译：用来自遗传丰富的小鼠细胞系的DNA制备了一个基因组文库，其中腺苷脱氨酶（ADA）基因的扩增拷贝占基因组的5％以上。从该基因组文库中分离出包含整个ADA结构基因的重叠粘粒克隆，并将其用于后续的结构和功能分析。核酸酶保护和引物延伸分析用于鉴定结构基因5'端多个转录起始位点的位置。通过功能分析发现启动子活性位于包含转录起始位点的240个碱基对的片段内。转录起始位点上游的序列非常富含G + C（77％），包括22个核苷酸的脱氧鸟苷残基延伸和两个潜在的Sp1转录因子结合位点。小鼠和人ADA基因启动子的比较揭示了在序列内容和位置方面高度保守的几个区域的存在，并且可能代表了参与ADA基因表达的遗传元件。

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