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哺乳动物G蛋白偶联受体的鉴定、染色体定位及基因组组织

Identification, chromosomal location, and genome organization of mammalian G-protein-coupled receptors.

作者信息

Wilkie T M, Chen Y, Gilbert D J, Moore K J, Yu L, Simon M I, Copeland N G, Jenkins N A

机构信息

Biology Division, California Institute of Technology, Pasadena 91125.

出版信息

Genomics. 1993 Nov;18(2):175-84. doi: 10.1006/geno.1993.1452.

Abstract

Vertebrate G-protein-coupled receptors are encoded by a diverse multigene family. Thirteen distinct G-protein-coupled receptors (Gpcr) were cloned from mouse germline cDNA following amplification in the polymerase chain reaction (PCR) with degenerate oligonucleotide primers complementary to the third and sixth transmembrane domains. Eleven Gpcr clones were mapped to single sites in the mouse genome following interspecific backcross analysis. One clone was mapped to two sites and another was not polymorphic in the cross and could not be mapped. Gpcr loci were well dispersed throughout the mouse genome and mapped to chromosomes 1, 2, 3, 4, 5, 8, 9, 10, 13, 17, and 18. Six Gpcr clones likely represent mouse homologs of already identified receptors, one Gpcr clone may identify a third type of IL-8 receptor, and three Gpcr clones appear to encode novel G-protein-coupled receptors. Further, three factor crosses and Southern blot analyses demonstrated that Gpcr16 maps proximally within the Spr deletion on mouse chromosome 1, near the Vil and Bcg loci. Human chromosomal locations for most Gpcr loci could be predicted based on linkage homologies that have been identified between human and mouse. Mapping additional G-protein-coupled receptors against the panel of murine interspecific backcrosses should expand our understanding of mammalian Gpcr gene evolution and genome organization.

摘要

脊椎动物的G蛋白偶联受体由一个多样的多基因家族编码。在用与第三和第六跨膜结构域互补的简并寡核苷酸引物进行聚合酶链反应(PCR)扩增后,从小鼠种系cDNA中克隆出13种不同的G蛋白偶联受体(Gpcr)。在种间回交分析后,11个Gpcr克隆被定位到小鼠基因组的单个位点。一个克隆被定位到两个位点,另一个在杂交中没有多态性,无法定位。Gpcr基因座在小鼠基因组中分布良好,定位于染色体1、2、3、4、5、8、9、10、13、17和18。6个Gpcr克隆可能代表已鉴定受体的小鼠同源物,1个Gpcr克隆可能鉴定出第三种类型的IL-8受体,3个Gpcr克隆似乎编码新的G蛋白偶联受体。此外,三因子杂交和Southern印迹分析表明,Gpcr16在小鼠1号染色体上Spr缺失的近端,靠近Vil和Bcg基因座。基于已确定的人与小鼠之间的连锁同源性,可以预测大多数Gpcr基因座的人类染色体位置。利用小鼠种间回交面板定位更多的G蛋白偶联受体,应该会扩展我们对哺乳动物Gpcr基因进化和基因组组织的理解。

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