Ano2是钙激活氯离子通道的分子基础

Ano2 is the molecular identity of calcium-activated chloride channels

目的探讨Ano2是否为钙激活氯离子通道的分子基础。方法用RT-PCR技术扩增Ano2编码基因,将Ano2连接到真核表达载体p EGFP-N3;通过脂质体介导将Ano2转染至FRT细胞,抗生素筛选获得稳定表达Ano2的细胞系;倒置荧光显微镜下观察Ano2在FRT细胞中的表达和分布,Western blot检测Ano2的表达;全细胞膜片钳技术研究Ano2的电生理学特性。结果成功构建p EGFP-Ano2真核表达载体;Ano2表达在FRT细胞膜上;Ano2电流呈Ca2+、时间和电压依赖性,电流和电压关系呈外向整流。结论 Ano2是钙激活氯离子通道的分子基础。

Objective To investigate that whether Ano2 is the molecular identity of calcium-activated chloride channels.Methods The full length coding sequence of Ano2 was amplified by RT-PCR,Ano2 and eukaryotic expression vector pEGFP-N3 were ligated;Then recombinant plasmids were transfected into FRT cells using liposome,and the stable transfection FRT cells were selected with antibiotic; the expression and location of Ano2 was observed by the inverted fluorescence microscope and its expression was detected by Western blot;the electrophysiological properties of Ano2 were researched by whole-cell patch-clamp technique.Results pEGFP-Ano2 was successfully constructed,FRT cell membrane expressed Ano2 ; the electrophysiological properties of Ano2 were Ca^2＋-,time-and voltage-dependent,and an outward-rectifying I/V relationship was observed.Conclusions Ano2 is the molecular identity of calcium-activated chloride channels.