O'Dowd B F, Nguyen T, Jung B P, Marchese A, Cheng R, Heng H H, Kolakowski L F, Lynch K R, George S R
Addiction Research Foundation, Toronto, Ontario, Canada.
Gene. 1997 Mar 10;187(1):75-81. doi: 10.1016/s0378-1119(96)00722-6.
We report the discovery of four novel human putative G-protein-coupled receptor (GPCR) genes. Gene GPR20 was isolated by amplifying genomic DNA with oligos based on the opioid and somatostatin related receptor genes and subsequent screening of a genomic library. Also, using our customized search procedure of a database of expressed sequence tags (dbEST), cDNA sequences that partially encoded novel GPCRs were identified. These cDNA fragments were obtained and used to screen a genomic library to isolate the full-length coding region of the genes. This resulted in the isolation of genes GPR21, GPR22 and GPR23. The four encoded receptors share significant identity to each other and to other members of the receptor family. Northern blot analysis revealed expression of GPR20 and GPR22 in several human brain regions while GPR20 expression was detected also in liver. Fluorescence in situ hybridization (FISH) was used to map GPR20 to chromosome 8q, region 24.3-24.2, GPR21 to chromosome 9, region q33, GPR22 to chromosome 7, region q22-q31.1, and GPR23 to chromosome X, region q13-q21.1.
我们报告发现了四个新的人类假定G蛋白偶联受体(GPCR)基因。基因GPR20是通过基于阿片样物质和生长抑素相关受体基因的寡核苷酸扩增基因组DNA,随后筛选基因组文库而分离得到的。此外,利用我们定制的表达序列标签数据库(dbEST)搜索程序,鉴定出了部分编码新型GPCR的cDNA序列。获得这些cDNA片段并用于筛选基因组文库,以分离基因的全长编码区。这导致了基因GPR21、GPR22和GPR23的分离。这四个编码的受体彼此之间以及与受体家族的其他成员具有显著的同源性。Northern印迹分析显示GPR20和GPR22在几个人脑区域表达,而在肝脏中也检测到GPR20的表达。荧光原位杂交(FISH)用于将GPR20定位到8号染色体q24.3 - 24.2区域,GPR21定位到9号染色体q33区域,GPR22定位到7号染色体q22 - q31.1区域,GPR23定位到X染色体q13 - q21.1区域。