摘要
目的Mipu1是在脑缺血预处理时表达上调的基因,本研究拟通过观察Mipu1蛋白在脑星形细胞瘤中的亚细胞定位,为进一步探讨其功能提供线索和思路。方法通过加端PCR扩增1.8 kb的Mipu1开放阅读框插入绿色荧光蛋白编码区的3’末端,构建Mipu1与绿色荧光蛋白GFP的融合表达质粒,并通过脂质体将重组质粒导入脑星形细胞瘤中,通过荧光显微镜观察Mipu1蛋白在正常和H2O2刺激时的亚细胞分布。结果成功构建了Mipu1与GFP的融合表达质粒Mipu1-pEGFP-N1,并观察到Mipu1蛋白在正常和H2O2刺激时均定位于细胞核内。结论Mipu1为一核蛋白,结合其蛋白质结构特征,推测其可能为一核转录因子。
Objective To investigate the subcellular localization of Mipul protein in brain astrocytoma cells and it＇s potential functional role. Methods The full-length open reading frame （ORF） of Mipul was fused to the 5＇ end of green fluorescent protein （GFP） coding sequence. Then the constructicn was transiently transfected into brain astrocytoma cells. The subcellular localization of Mipul was analyzed by fluorescence microscope before and after the cells were stimulated by H2O2. Results The Mipul-GFP fusion construction was successfully transfected into brain astrocytoma ceils. Fluorescence microscopy demonstrated that Mipu1 was located in cellular nucleus under normal condition. However there was no change of subcellular localization of Mipul when the cells were stimulated by H2O2. Conclusion Mipul is comnsidered as a nuclear transcription factor based on it＇s protein structure and subcellular localization.
出处
《中国耳鼻咽喉颅底外科杂志》
CAS
2006年第3期164-167,共4页
Chinese Journal of Otorhinolaryngology-skull Base Surgery
