作　　者： ; ; ; ;

机构地区： 南方医科大学生物技术学院生物治疗研究所

出　　处： 《中国细胞生物学学报》 2010年第3期461-464,共4页

摘　　要： 采用RT-PCR的方法扩增人Mcm6基因,克隆入真核表达载体pEGFP-N3,成功构建了pEGFP-Mcm6真核表达质粒,酶切及DNA测序证实了构建质粒序列的正确性。脂质体介导转染Hela细胞后,荧光显微镜可检测到绿色荧光信号,通过G418筛选最终获得了稳定表达的细胞株,Western blot验证了稳定表达细胞株中Mcm6-GFP融合蛋白的表达。本实验成功建立了稳定表达Mcm6-GFP融合蛋白的细胞株,为动态观察Mcm6细胞定位的变化,进一步研究抗肿瘤药物的作用机制提供了有效的分子工具。 Human Mcm6 gene was amplified by RT-PCR and cloned into pEGFP-N3 vector to get pEGFP- Mcm6 plasmid. The recombinant plasmid was confirmed by enzyme digestion and DNA sequencing. After transfection into Hela cells, green signals were observed under fluorescence microscopy. Under pressure of G418, stable cell lines were appeared and Western blot confirmed the expression of Mcm6-GFP fusion protein. In this study, stable cell line expressing Mcm6-GFP fusion protein was successfully established, by which the dynamic localization of Mcm6 can be studied.

关 键 词： 融合蛋白 稳定表达

领　　域： [生物学]