Itoh S, Abe Y, Kubo A, Okuda M, Shimoji M, Nakayama K, Kamataki T
Division of Drug Metabolism, Faculty of Pharmaceutical Sciences, Hokkaido University, Japan.
Biochim Biophys Acta. 1997 Feb 7;1350(2):155-8. doi: 10.1016/s0167-4781(96)00215-1.
An 11.5 kb fragment of the mouse Cyp3a16 gene containing the 5' flanking region was isolated from the lambda DASHII mouse genomic library. A part of the 5' flanking region and the first exon of Cyp3a16 gene were sequenced. S1 mapping analysis showed the presence of two transcriptional initiation sites. The first exon was completely identical to Cyp3a16 cDNA. The identity of 5' flanking sequences between Cyp3a16 and Cyp3a11 genes was about 69%. A typical TATA box and a basic transcription element (BTE) were found as seen with other CYP3A genes from various animal species Moreover, some putative transcriptional regulatory elements were also found in addition to the sequence motif seen for the formation of Z-type DNA. To examine the transcriptional activity of Cyp3a11 gene, DNA fragments in the 5'-flanking region of the gene were inserted front of the luciferase structural gene, and the constructs were transfected in primary hepatocytes. The analysis of the luciferase activity indicated that the region between -146 and -56 was necessary for the transcription of CYP3a16 gene.
从小鼠λDASHII基因组文库中分离出包含5'侧翼区域的11.5 kb小鼠Cyp3a16基因片段。对Cyp3a16基因5'侧翼区域的一部分和第一个外显子进行了测序。S1图谱分析显示存在两个转录起始位点。第一个外显子与Cyp3a16 cDNA完全相同。Cyp3a16和Cyp3a11基因之间5'侧翼序列的同一性约为69%。与来自各种动物物种的其他CYP3A基因一样，发现了一个典型的TATA盒和一个基本转录元件（BTE）。此外，除了Z型DNA形成所观察到的序列基序外，还发现了一些推定的转录调控元件。为了检测Cyp3a11基因的转录活性，将该基因5'侧翼区域的DNA片段插入荧光素酶结构基因的前面，并将构建体转染到原代肝细胞中。荧光素酶活性分析表明，-146至-56之间的区域对于CYP3a16基因的转录是必需的。
