带绿色荧光蛋白标签的人era真核表达载体的构建及人Era对细胞形态和细胞周期的影响  

Construction of human era eukaryotic expression vector with EGFP tag and effects of human Era on cell morphogenesis and cell cycle

作　　者：纪宗玲[1] 陈苏民[1] 刘断中[2] 吴元明[1] 张俊杰[1] 路凡[1] 朱帮福[1] 李毅[1] 

机构地区：[1]第四军医大学生物化学与分子生物学教研室,陕西西安710032 [2]第四军医大学西京医院全军骨科研究所,陕西西安710032

出　　处：《中国病理生理杂志》2003年第11期1445-1448,U002,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目 (No .39870 380;30 0 0 0 0 91) ;全军医药卫生科研基金资助项目 (98M10 8)

摘　　要：目的 :构建人era真核表达载体并初步研究这一人类新基因对体外培养细胞形态和细胞周期的作用。方法 :构建带绿色荧光蛋白 (EGFP)标签的人era真核表达载体 ,转染体外培养细胞NIH3T3,用荧光显微镜观测和流式细胞仪进行分析。结果 :无论EGFP融合于人Era的C端或N端 ,融合蛋白均表达于细胞浆接近核膜处 ,细胞形态无明显变化 ,细胞周期检测S期细胞所占百分数降低。结论 :人era可能参与细胞周期调控 ,为阐明人era的功能提供了资料。AIM: To construct eukaryotic expressive vectors of human era and analyze the effect of era on morphogenesis and cell cycle in vitro . METHODS: Two eukaryotic expressive vectors of EGFP-hEra fusion protein named pSMEGFP-hEra and pEGFP-C1-hEra were constructed and transfected into mouse fibroblast cell line NIH3T3 by way of lipofectin. Morphogenesis and cell cycle were analyzed by fluorescence microscopy and flow cytometer,respectively. RESULTS: Fusion protein of EGFP and human Era localized in cytoplasm around cell nucleus mainly. No morphologic change was examined,cell cycle analysis revealed the cell number of S phase lightly decreased. CONCLUSION: Human era may play a key role in cell cycle,these results may shed light on the further study of the function of this new gene.

关 键 词：基因表达 细胞形态 细胞周期 人era真核表达载体 绿色荧光蛋白 ERA基因 

分 类 号：Q987[生物学—遗传学]