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G蛋白偶联受体家族四个新成员的选择性扩增与克隆

Selective amplification and cloning of four new members of the G protein-coupled receptor family.

作者信息

Libert F, Parmentier M, Lefort A, Dinsart C, Van Sande J, Maenhaut C, Simons M J, Dumont J E, Vassart G

机构信息

Institut de Recherche Interdisciplinaire, Faculté de Médecine, Université Libre de Bruxelles, Campus Erasme, Belgium.

出版信息

Science. 1989 May 5;244(4904):569-72. doi: 10.1126/science.2541503.

Abstract

An approach based on the polymerase chain reaction has been devised to clone new members of the family of genes encoding guanosine triphosphate-binding protein (G protein)-coupled receptors. Degenerate primers corresponding to consensus sequences of the third and sixth transmembrane segments of available receptors were used to selectively amplify and clone members of this gene family from thyroid complementary DNA. Clones encoding three known receptors and four new putative receptors were obtained. Sequence comparisons established that the new genes belong to the G protein-coupled receptor family. Close structural similarity was observed between one of the putative receptors and the 5HT1a receptor. Two other molecules displayed common sequence characteristics, suggesting that they are members of a new subfamily of receptors with a very short nonglycosylated (extracellular) amino-terminal extension.

摘要

已设计出一种基于聚合酶链反应的方法来克隆编码鸟苷三磷酸结合蛋白(G蛋白)偶联受体的基因家族的新成员。对应于现有受体第三和第六跨膜区段共有序列的简并引物用于从甲状腺互补DNA中选择性扩增和克隆该基因家族的成员。获得了编码三种已知受体和四种新的推定受体的克隆。序列比较确定这些新基因属于G蛋白偶联受体家族。在一种推定受体与5HT1a受体之间观察到紧密的结构相似性。另外两个分子显示出共同的序列特征,表明它们是具有非常短的非糖基化(细胞外)氨基末端延伸的受体新亚家族的成员。

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