Cloning and expression of an A sub(1) adenosine receptor from rat brain

• Published in: Molecular pharmacology Vol. 40; no. 1; pp. 1 - 7
• Main Authors: Mahan, L C, Mcvittie, L D, Smyk-Randall, E M, Nakata, H, Monsma, FJ Jr, Gerfen, C R, Sibley, DR
• Format: Journal Article
• Language: English
• Published: 01.01.1991
• Subjects: brain; genes; mRNA
• ISSN: 0026-895X

We have used the polymerase chain reaction technique to selectively amplify guanine nucleotide-binding regulatory protein (G protein)-coupled receptor cDNA sequences from rat striatal mRNA, using sets of highly degenerate primers derived from transmembrane sequences of previously cloned G protein-coupled receptors. A novel cDNA fragment was identified, which exhibits considerable homology to various members of the G protein-coupled receptor family. This fragment was used to isolate a full-length cDNA from a rat striatal library. A 2.2-kilobase clone was obtained that encodes a protein of 326 amino acids with seven transmembrane domains, as predicted by hydropathy analysis. Receptor mRNA in brain tissues revealed two transcripts of 5.6 and 3.1 kilobases, both of which were abundant in cortex, cerebellum, hippocampus, and thalamus, with lower levels in olfactory bulb, striatum, mesencephalon, and retina. These regional distribution data are in good agreement with previous receptor autoradiographic studies involving the A sub(1) adenosine receptor.