Cloning of two additional catecholamine receptors from rat brain

FEBS Letters

Jarvie, Keith R.¹  Israel, Yedy²  Tirpak, Anne¹  Niznik, Hyman B.¹  Seeman, Philip¹  Nguyen, Tuan²  O'Dowd, Brian F.² 

[1] Departments of Pharmacology, University of Toronto, Toronto, Ont.Canada M5S 1A8;Addiction Research Foundation, 33 Russell Street, Toronto, Ont.Canada M5S 2S1

关键词: Polymerase chain reaction;    Homology probing;    D2 dopamine receptor;    Receptor variant;    AR;    adrenergic receptor;    PCR;    polymerase chain reaction;    TM;    transmembrane;    G;    guanine nucleotide binding protein;   

DOI  :  10.1016/0014-5793(90)80140-E

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

An approach based on the polymerase chain reaction (PCR) was used to isolate additional members of the G-linked receptor family from a rat striatal λgtII cDNA library. Priming with one degenerate probe corresponding to highly conserved consensus sequences in the third transmembrane (TM) domain of 15 G-linked receptors and sequences in the phage vector resulted in one clone (G-13) encoding a dopamine D2 receptor variant with a 29 amino acid insert in the third cytoplasmic loop. In addition, the amino acid sequence encoded by clone G-36 contained conserved sequences characteristic of the G-linked class of receptors and displayed sequence homology in TM domains with the β₂-adrenergic receptor (48%). Two conserved serine residues in TM5 postulated to be part of a ligand binding site in the adrenergic receptor, suggests that G-36 encodes a catecholaminergic receptor. Northern blot analysis confirmed the expression of G-36 in rat brain, but not in kidney, heart and lung. Several strong hybridizing bands to G-36 were obtained in both human and rat genomic DNA. The general PCR strategy employed here should prove to be extremely useful for the isolation of other members of the G-linked receptor family.

【 授权许可】

Unknown