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一种人ATP调节钾通道的分离与染色体定位

Isolation and chromosomal localization of a human ATP-regulated potassium channel.

作者信息

Krishnan S N, Desai T, Ward D C, Haddad G G

机构信息

Department of Pediatrics (Section of Respiratory Medicine), Yale University School of Medicine, New Haven, CT 06520-8064, USA.

出版信息

Hum Genet. 1995 Aug;96(2):155-60. doi: 10.1007/BF00207372.

Abstract

KATP channels are K+ channels whose activity is inhibited by the presence of and enhanced by the absence of cytosolic ATP. This property allows KATP channels to sense cellular intermediary metabolism and directly couple this information to the modulation of membrane excitability. Indeed, recent studies from our laboratory and others have suggested that activation of KATP channels during anoxia is important in the response and adaptation of central neurons to hypoxia. In order to identify KATP channels from human brain, we performed a polymerase chain reaction (PCR) using human cerebral cortex mRNA and primers derived from the ROMK1 sequence, a cDNA clone encoding an ATP-regulated potassium channel, recently isolated from rat kidney. We thus identified a novel 308-bp PCR product, pKCNJ1, whose expression was found to be restricted to a 3.0-kb band in the kidney by probing a human multiple tissue northern blot, pKCNJ1 was then used to isolate genomic clones and, using fluorescence in situ hybridization (FISH) to human metaphase chromosomes, was mapped to chromosome 11q.

摘要

KATP通道是一种钾离子通道,其活性在胞质ATP存在时受到抑制,而在胞质ATP缺失时增强。这一特性使KATP通道能够感知细胞中间代谢,并将该信息直接与膜兴奋性的调节相联系。事实上,我们实验室及其他机构最近的研究表明,缺氧时KATP通道的激活对于中枢神经元对缺氧的反应和适应至关重要。为了从人脑中鉴定KATP通道,我们使用人大脑皮质mRNA和源自ROMK1序列的引物进行了聚合酶链反应(PCR),ROMK1是一个编码ATP调节钾通道的cDNA克隆,最近从大鼠肾脏中分离得到。我们由此鉴定出一种新的308 bp PCR产物pKCNJ1,通过探测人多组织Northern印迹发现其表达仅限于肾脏中的一条3.0 kb条带。然后用pKCNJ1分离基因组克隆,并利用荧光原位杂交(FISH)技术将其定位到人类中期染色体上,结果显示它位于11号染色体长臂(11q)。

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