cDNA cloning of a putative G protein-coupled receptor from brain.
Using degenerate oligonucleotide primers corresponding to conserved regions of the G-protein coupled receptor superfamily, we carried out a low-stringency polymerase chain reaction and obtained two novel partial-length clones from a rat brain cDNA library. We used one of these clones for conventional library screening and isolated a longer cDNA clone, designated as RBU-15, from another rat brain library. Although RBU-15 was truncated at its 5' end, Northern blot analysis revealed that the gene was expressed in the brain and spleen. Next, we isolated a full-length cDNA clone, designated as HB-954, from a human fetal brain library, using RBU-15 as a probe. The deduced amino acid sequence of HB-954 contained four putative glycosylation sites in the N-terminal part, seven transmembrane domains, and a large cytosolic domain in the C-terminal part. The protein products of RBU-15 and HB-954 likely belong to a distinctive subfamily, because no receptors in the superfamily were found to be closely related to them.
Hata
S
S
Department of Life Science, Faculty of Science, Himeji Institute of Technology, Hyogo, Japan.
Emi
Y
Y
Iyanagi
T
T
Osumi
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001923:Brain Chemistry; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D019204:GTP-Binding Proteins / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D005810:Multigene Family; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000302:isolation & purification; D010802:Phylogeny; D051381:Rats; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000302:isolation & purification; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D018076:DNA, Complementary; D009419:Nerve Tissue Proteins; D011956:Receptors, Cell Surface; D019204:GTP-Binding Proteins