Bruno J F, Whittaker J, Song J F, Berelowitz M
Department of Medicine, State University of New York, Stony Brook 11794.
Biochem Biophys Res Commun. 1991 Sep 30;179(3):1485-90. doi: 10.1016/0006-291x(91)91740-4.
DNA from a rat hippocampus cDNA library and sets of highly degenerate oligonucleotide primers directed toward conserved regions of previously cloned G-protein receptors were used in the polymerase chain reaction to selectively amplify and clone new members of this gene family. A human hippocampus cDNA library was screened with a 610 base pair fragment generated by PCR and a cDNA clone, H318/3, was isolated. The deduced amino acid sequence of this clone encoded a protein of 501 amino acids that showed strong sequence homology to previously cloned G-protein receptors. Nucleotide sequence analysis revealed clone H318/3 was 78% homologous to a rat alpha 1A adrenergic receptor with homology being 95% when comparisons were made in the region that lies between the first to the seventh transmembrane domains. Based on this high degree of sequence homology, we conclude that clone H318/3 represents a cDNA for a human alpha 1A adrenergic receptor.
利用大鼠海马cDNA文库的DNA以及针对先前克隆的G蛋白受体保守区域设计的高度简并寡核苷酸引物，通过聚合酶链反应选择性扩增并克隆该基因家族的新成员。用PCR产生的610个碱基对的片段筛选人海马cDNA文库，分离出一个cDNA克隆H318/3。该克隆推导的氨基酸序列编码一个由501个氨基酸组成的蛋白质，该蛋白质与先前克隆的G蛋白受体具有很强的序列同源性。核苷酸序列分析显示，克隆H318/3与大鼠α1A肾上腺素能受体的同源性为78%，在第一至第七跨膜结构域之间的区域进行比较时，同源性为95%。基于这种高度的序列同源性，我们得出结论，克隆H318/3代表人类α1A肾上腺素能受体的cDNA。
