Cloning of two additional catecholamine receptors from rat brain.
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 lambda gtII 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 beta 2-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.
O'Dowd
B F
BF
Addiction Research Foundation, Toronto, Ont. Canada.
Nguyen
T
T
Tirpak
A
A
Jarvie
K R
KR
Israel
Y
Y
Seeman
P
P
Niznik
H B
HB
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003342:Corpus Striatum / Q000032:analysis; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D051381:Rats; D011954:Receptors, Dopamine / Q000032:analysis / Q000235:genetics*; D017448:Receptors, Dopamine D2
D011954:Receptors, Dopamine; D017448:Receptors, Dopamine D2