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cDNA cloning of novel seven-transmembrane receptors from rat brain

大鼠脑中新型七次跨膜受体的 cDNA 克隆

基本信息

批准号：
05680682
负责人：
HATA Shingo
金额：
$ 1.34万
依托单位：
KYOTO UNIVERSITY (1994)Himeji Institute of Technology (1993)
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

项目摘要

Using degenerate oligonucleotide primers corresponding to conserved regions of the G-protein coupled receptor (seven-transmembrane receptor) superfamily, we carried out a low-stringency polymerase chain reaction and obtained two novel partial-length clones from a rat brain cDNA library. We used one of these clones for conventional library screening and isolated a longer cDNA clone, designated as RBU-15, from another rat brain library. Although RBU-15 was truncated at its 5' end, Northern blot analysis revealed that the gene was expressed in the brain and spleen. Next, we isolated a full-length cDNA clone, designated as HB-954, from a human fetal brain library, using RBU-15 as a probe. The deduced amino acid sequence of HB-954 contained four putative glycosylation sites in the N-terminal part, seven transmembrane domains, and a large cytosolic domain in the C-terminal part. The protein products of RBU-15 and HB-954 likely belong to a distinctive subfamily, because no receptors in the superfamily were found to be closely related to them.

使用简并寡核苷酸引物对应的G蛋白偶联受体（七跨膜受体）超家族的保守区域，我们进行了低严格的聚合酶链反应，并获得了两个新的部分长度克隆从大鼠脑cDNA文库。我们使用这些克隆之一进行常规文库筛选，并从另一个大鼠脑文库中分离出一个更长的cDNA克隆，命名为RBU-15。尽管RBU-15的5'端被截短，但北方印迹分析表明该基因在脑和脾中表达。接下来，我们用RBU-15作为探针，从人胎脑文库中分离出一个全长cDNA克隆，命名为HB-954。推导的HB-954氨基酸序列在N-末端部分含有4个推测的糖基化位点，7个跨膜结构域，在C-末端部分含有一个大的胞质结构域。RBU-15和HB-954的蛋白产物可能属于一个独特的亚家族，因为在超家族中没有发现与它们密切相关的受体。