CAJ | 학술논문

目的建立稳定表达TMEM16A蛋白的TE-1肿瘤细胞株.方法用Lipofectamine?2000转染试剂将TMEM16A基因转染到TE-1细胞,经G418筛选,使用激光共聚焦显微镜和Western blot技术检测TMEM16A基因是否在TE-1细胞中已经稳定表达.结果通过使用G418进行筛选,阳性克隆于4周后培养形成.激光共聚焦显微镜观察稳定表达TMEM16A蛋白的TE-1细胞可见细胞发出绿色荧光.Western blot结果表明,未稳转细胞和稳转细胞系统均表达有TMEM16A蛋白,而稳转细胞系统的蛋白水平明显高于未转染细胞系统的蛋白水平.结论利用脂质体转染方法成功构建了稳定表达TMEM16A基因的TE-1细胞系.
목적건립은정표체TMEM16A단백적TE-1종류세포주.방법용Lipofectamine?2000전염시제장TMEM16A기인전염도TE-1세포,경G418사선,사용격광공취초현미경화Western blot기술검측TMEM16A기인시부재TE-1세포중이경은정표체.결과통과사용G418진행사선,양성극륭우4주후배양형성.격광공취초현미경관찰은정표체TMEM16A단백적TE-1세포가견세포발출록색형광.Western blot결과표명,미은전세포화은전세포계통균표체유TMEM16A단백,이은전세포계통적단백수평명현고우미전염세포계통적단백수평.결론이용지질체전염방법성공구건료은정표체TMEM16A기인적TE-1세포계.
Objective To get stably expressing TMEM16A proteins by transfecting TE-1 cells. Methods TMEM16A gene was transfected into TE-1 cells by Lipofectamine?2000, then we picked up cell clones in the presence of G418. TMEM16A proteins were identified by confocal microscope and western blot. Results After 4 weeks, the cell clones of colony-like formed after G418 screening. The stable TMEM16A-expression TE-1 cell lines were observed by confocal microscope showing signal of GFP co-transfected. Western blot results showed that TMEM16A protein expressed in control and stable TMEM16A-expression TE-1 cells, and the expression was significantly increased in TMEM16A stable cell lines. Conclusion Stable TMEM16A-expression TE-1 cell lines are created with liposome method successfully.