Morishige K, Takahashi N, Findlay I, Koyama H, Zanelli J S, Peterson C, Jenkins N A, Copeland N G, Mori N, Kurachi Y
Department of Internal Medicine, Mayo Clinic, Rochester, MN 55905.
FEBS Lett. 1993 Dec 28;336(3):375-80. doi: 10.1016/0014-5793(93)80840-q.
We have cloned an inward-rectifier potassium channel from a mouse brain cDNA library, studied its distribution in the brain by in situ hybridization and determined the chromosomal localization of the gene. A mouse brain cDNA library was screened using a fragment of the mouse macrophage IRK1 cDNA as a probe. Two duplicate clones of approximately 5.5 kb were obtained. Xenopus ococytes injected with cRNA derived from the clone expressed a potassium channel with inwardly rectifying channel characteristics. The amino acid sequence of the clone was identical to that of IRK1 recently cloned from a mouse macrophage cell line. In situ hybridization study showed the mouse brain IRK1 to be generally distributed throughout the brain, but in particular subsets of neurons at high levels. The gene was placed in the distal region of mouse chromosome 11, which contains several uncloned neurological mutations. These results provide the first demonstration of the cloning and distribution of an inward rectifier potassium channel from the nervous system.
我们从小鼠脑cDNA文库中克隆了一种内向整流钾通道,通过原位杂交研究了其在脑中的分布,并确定了该基因的染色体定位。以小鼠巨噬细胞IRK1 cDNA片段为探针筛选小鼠脑cDNA文库。获得了两个约5.5 kb的重复克隆。注射来自该克隆的cRNA的非洲爪蟾卵母细胞表达出具有内向整流通道特性的钾通道。该克隆的氨基酸序列与最近从小鼠巨噬细胞系克隆的IRK1相同。原位杂交研究表明,小鼠脑IRK1普遍分布于整个脑,但在特定的神经元亚群中含量较高。该基因位于小鼠11号染色体的远端区域,该区域包含几个未克隆的神经学突变。这些结果首次证明了从神经系统克隆和分布内向整流钾通道。