中心体蛋白centrin2稳定高表达慢性髓原白血病细胞系的构建及鉴定

Construction and Identification of Centrosome Protein Centrin2 Over-expression Chronic Myeloid Leukemia Cell Lines

将质粒转染慢性髓原白血病细胞K562,经过G418筛选获得稳定高表达centrin2的细胞株.通过免疫荧光显微镜观察细胞的变化,使用蛋白免疫印迹方法鉴定细胞株内centrin2蛋白的过表达效果,并通过细胞免疫荧光染色技术方法观察中心体上centrin2含量的改变.成功获得了一株稳定高表达centrin2的K562细胞株(K562-GFP-centrin2)和稳定表达无意义GFP的对照细胞(K562-GFP).成功构建了稳定高表达centrin2蛋白的慢性髓原白血病细胞K562-GFP-centrin2,为细胞中心体成功的提取奠定基础.

Expession plasmids of pGFP - centrin2 and pGFP were transfected into chronic myeloid leukemia cells （K562）. The cells were cultured in selection medium containing G418 to form single clones. Finally, stable centrin2 over - expressing K562 cells were picked up under an fluoroscent microscope and identified by Western Blot. Two cell lines stably over - expressing pGFP - centrin2 and one with pGFP were successfully constructed. Stable over- expressing pGFP- centrin2 K562 cell lines were successfully constructed that can serve for further study of extraction of eentrosome.