Cammas F M, Kapas S, Barker S, Clark A J
Department of Chemical Endocrinology, St Bartholomew's Hospital Medical College, London, United Kingdom.
Biochem Biophys Res Commun. 1995 Jul 26;212(3):912-8. doi: 10.1006/bbrc.1995.2056.
A polymerase chain reaction-generated mouse ACTH-R probe was used to screen a mouse genomic library, and a clone of 13kb containing the entire coding sequence was isolated. The coding sequence shows 84% homology with the human gene at the DNA level and encodes a peptide with 89% homology to the human ACTH-R. This gene is expressed as a major transcript of 1.8kb in the mouse adrenal gland. The gene was expressed in HeLa cells and cAMP production in response to either ACTH or alpha-MSH was measured. cAMP increased in an ACTH dose dependent manner suggesting an EC50 of 7 x 10(-10)M ACTH. alpha-MSH was without effect on this receptor. In conclusion we have cloned a mouse ACTH receptor gene and demonstrated for the first time its expression and functional effect in HeLa cells.
使用聚合酶链反应产生的小鼠促肾上腺皮质激素受体（ACTH-R）探针筛选小鼠基因组文库，分离出一个包含完整编码序列的13kb克隆。该编码序列在DNA水平上与人基因显示出84%的同源性，并编码一种与人类促肾上腺皮质激素受体有89%同源性的肽。该基因在小鼠肾上腺中表达为1.8kb的主要转录本。该基因在HeLa细胞中表达，并检测了其对促肾上腺皮质激素（ACTH）或α-促黑素细胞激素（α-MSH）的反应中cAMP的产生。cAMP以ACTH剂量依赖性方式增加，表明ACTH的半数有效浓度（EC50）为7×10^(-10)M。α-MSH对该受体无作用。总之，我们克隆了小鼠促肾上腺皮质激素受体基因，并首次证明了其在HeLa细胞中的表达和功能作用。
