不同剪切形式的小鼠Era对L-929细胞的影响

Effects of different splicing mouse Era on L- 929 cells

目的: 构建两种完全可调控诱导不同剪接形式小鼠era的真核表达载体,检测不同剪接形式的鼠Era蛋白对细胞生长状态的影响.方法: 构建两种不同剪接形式鼠era的完全可调控诱导表达载体,分别与受体表达载体pERV3稳定共转染鼠成纤维细胞系L 929.用PonA进行诱导表达后,以West ernBlotting检测Era蛋白表达水平,以MTT法测定细胞生长曲线,以流式细胞仪检测细胞周期.结果: 成功获得稳定的可完全诱导表达两种不同剪接形式鼠Era蛋白的细胞系;经过PonA诱导后,过表达两种剪接形式Era蛋白的L 929细胞,生长速度加快,G2 /M期细胞数量明显减少.结论: 两种剪接形式鼠Era蛋白在真核细胞周期中发挥重要作用.

AIM: To construct complete inducible mammalian expression vectors that can express two alternative splicing mouse eras and to study the effects of two alternative splicing mouse era genes on cells growth.METHODS: Two inducible vectors that express two alternative splicing mouse Eras were constructed.The inducible vectors and pERV3 were then stably co-transfected into L-929 cells.After transfected cells were induced by PonA,the protein expression was detected by Western Blotting,the proliferation of cells was measured by MTT and the cell cycles were observed by flow cytometry.RESULTS: Two kinds of cell lines that can express different splicing mouse Era proteins were obtained.The growth of cells was accelerated when the cells were induced by PonA and the percentages of G2/M phase cells decreased.CONCLUSION: Two alternative splicing genes play important roles in the eukaryote cell cycle.