Cloning and characterization of a new member of the G-protein coupled receptor EDG family.

摘要: We report here the cloning of a new member endothelium differentiation gene (edg) subfamily G-protein-coupled receptors. This novel cDNA sequence was cloned from ovine pars tuberalis using reverse transcriptase polymerase chain reaction (RT-PCR) amplification with degenerate primers homologous to highly conserved II and VII transmembrane domains G-protein coupled receptor family. The PCR product random primed 32P used as probe screen size-selected library, which resulted in isolation single clone 2700 bp. named edg-2, because its nucleic acid 55% over 501nt overlap an orphan human endothelial cells, gene, edg-1. highest degree aminoacid homology (42%) occurs seven putative domains, particularly between III VI (53% 64%, respectively). intervening hydrophilic are short there numerous phosphorylation sites for Ser/Thr-protein kinases second third intracellular loop COOH-terminal domain. Through Northern analysis total RNA, low levels at least four transcripts 2.3, 2.5, 3.2 4 kb were found sheep cerebral cortex 4.2 transcript observed NIH/3T3 fibroblasts. In addition, edg-2 (415 bp) amplified by RT-PCR tuberalis, blood vessels, hypothalamus, retina. Serum stimulation Chinese hamster ovary (CHO) cells expressing increased cell proliferation, measured [3H]-thymidine incorporation. Edg-1 appear be distinct genes that may encode protein products bind same or related ligand.