Janssens R, Communi D, Pirotton S, Samson M, Parmentier M, Boeynaems J M
Institute of Interdisciplinary Research, School of Medicine, Erasme Hospital, Université Libre de Bruxelles, Belgium.
Biochem Biophys Res Commun. 1996 Apr 25;221(3):588-93. doi: 10.1006/bbrc.1996.0640.
Sets of degenerate oligonucleotide primers synthesized on the basis of the best conserved regions of the chick brain P2Y/P2Y1 and the murine neuroblastoma P2U/P2Y2 receptors were used in polymerase chain reaction experiments on human genomic DNA. An amplified fragment of 712 base pairs was then used as a probe to screen a human genomic DNA library. Several clones were isolated and sequencing revealed an intronless 1122 base pair open reading frame. The corresponding amino acid sequence revealed 83% identity with the chick brain P2Y1 receptor and 34% with the murine neuroblastoma P2Y2 receptor. In COS-7 cells transfected with the coding sequence inserted into the pcDNA3 expression vector, 2-methylthioATP and ATP produced a strong stimulation of inositol phosphates, a typical response of a P2Y1 receptor. Northern blot analysis detected a 6.7 kilobase messenger RNA in most human tissues, the strongest signals being observed in prostate and ovary.
根据鸡脑P2Y/P2Y1和鼠神经母细胞瘤P2U/P2Y2受体的最佳保守区域合成的简并寡核苷酸引物组，用于对人类基因组DNA进行聚合酶链反应实验。然后，将一个712个碱基对的扩增片段用作探针，筛选人类基因组DNA文库。分离出了几个克隆，测序显示有一个无内含子的1122个碱基对的开放阅读框。相应的氨基酸序列与鸡脑P2Y1受体有83%的同一性，与鼠神经母细胞瘤P2Y2受体有34%的同一性。在用插入到pcDNA3表达载体中的编码序列转染的COS-7细胞中，2-甲硫基ATP和ATP对肌醇磷酸产生了强烈的刺激作用，这是P2Y1受体的典型反应。Northern印迹分析在大多数人类组织中检测到一个6.7千碱基的信使RNA，在前列腺和卵巢中观察到最强的信号。
