【摘要】 为研究核基质结合区结合蛋白的功能及调控机制,PCR扩增杜氏盐藻MBP的cDNA全长序列及N端和C端序列,与绿色荧光蛋白基因融合构建真核表达载体,脂质体转染CHO细胞,Western blotting和荧光显微镜检测基因表达情况和细胞定位。结果显示:MBP及N端和C端融合蛋白成功在CHO细胞表达,MBP和C端部分定位于细胞核且聚集于核仁,N端部分分布整个细胞,说明MBP定位于细胞核且细胞定位信号位于C端,MBP可能与rRNA前体结合发挥作用。更多还原

【Abstract】 To study the function of Matrix attachment region(MAR)binding protein(MBP),full length,N-terminal and C-terminal of MBP gene from Dunaliella salina were amplified by RT-PCR,and fused to the green fluorescence protein gene to construct the eukaryotic vector.Then,the recombinant vectors were transfected into Chinese hamster ovary(CHO) cells by Lipofectamine 2000.Fluorescence microscope and western blotting were performed to determine the expression of the fusion proteins and cellular localization.The results showed that the fusion proteins were expressed in CHO cells,the proteins of MBP and C-terminal part were localized in the nucleus and clearly detected in nucleoli,however,the N-terminal part was distributed into all cells.更多还原