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胞质型磷脂酶A2(cPLA2)启动子的分离

Isolation of promoter for cytosolic phospholipase A2 (cPLA2).

作者信息

Tay A, Maxwell P, Li Z, Goldberg H, Skorecki K

机构信息

Division of Nephrology, University of Toronto, Canada.

出版信息

Biochim Biophys Acta. 1994 Apr 6;1217(3):345-7. doi: 10.1016/0167-4781(94)90299-2.

Abstract

Cytosolic phospholipase A2 (cPLA2) releases arachidonic acid from membrane phospholipids and is believed to be the rate-limiting enzyme in the arachidonic acid pathway. We report herein the isolation of a 3 kb fragment of rodent genomic DNA containing part of the first intron, the first exon and 5'-flanking sequence. The start site of transcription was mapped by 5'-rapid amplification of cDNA ends and corroborated by ribonuclease protection assay. The gene has a TATAless promoter with no classical Sp1 binding sites or initiator element. A microsatellite series of CA repeats was noted in the 5'-flanking region of both the rodent and human promoters. Deletion constructs have been analysed for luciferase activity and confirmed promoter activity.

摘要

胞质型磷脂酶A2(cPLA2)从膜磷脂中释放花生四烯酸,被认为是花生四烯酸途径中的限速酶。我们在此报告了从啮齿动物基因组DNA中分离出一个3 kb的片段,该片段包含部分第一内含子、第一外显子和5'侧翼序列。通过5'-cDNA末端快速扩增确定了转录起始位点,并通过核糖核酸酶保护试验得到了证实。该基因有一个无TATA盒的启动子,没有经典的Sp1结合位点或起始子元件。在啮齿动物和人类启动子的5'侧翼区域都发现了一系列CA重复的微卫星序列。已对缺失构建体进行了荧光素酶活性分析,并证实了启动子活性。

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