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

作　　者：藏雨轩[1] 方芳[2] 朱杭飞 向国艳[1] 张雲乔 李瑷彤 郝峰[1] 

机构地区：[1]吉林医药学院生物化学检验教研室,吉林吉林132013 [2]吉林医药学院免疫学教研室,吉林吉林132013

出　　处：《基础医学与临床》2014年第11期1477-1481,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(81202031);吉林省教育厅"十二五"科学技术研究项目(2013-351);2013年吉林省大学生创新创业训练计划(856号);吉林医药学院大学生科研项目[吉医学科字(2012)第12号]

摘　　要：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.

关 键 词：Ano2 FRT细胞 转染 钙激活氯离子通道 

分 类 号：Q249[生物学—细胞生物学]