钙结合蛋白CIB1在293T细胞中的表达及亚细胞定位研究

分析和比对钙结合蛋白CIB1在人类、大鼠、小鼠中氨基酸序列差异,并研究CIB1蛋白在293T细胞中的表达及亚细胞定位情况。通过Western Blot分析发现293T细胞本身几乎检测不到CIB1的表达。进一步采用CIB1外源性质粒转染,通过免疫荧光分析实验,在激光共聚焦显微镜下观察转染后不同时间293T细胞中CIB1的表达情况及亚细胞定位变化。实验结果表明,随着转染时间的增加,CIB1在293T细胞中的表达有逐渐增强的趋势,并且发生从细胞核向细胞质的转位现象。该实验结果对了解CIB1亚细胞定位变化及功能研究具有重要参考价值。

A Study on the Expression and Subcellular Localization of Calcium and Integrin Binding Protein 1 in 293 T Cells

Alignment of CIB1 protein sequence among human,rat and mouse,and study on the expression and subcul-luar localization of CIB1 in 293T cells. Western blot analysis show that CIB1 protein expression is increased significantly in transfected 293T cells, but undetectable in untransfected 293T cells. To further monitor the expression and subcellu-lar localization of CIB1 in 293T cells,transfections CIB1 exogenous plasmid transfection with varying time were detected by immunofluorescence analysis in laser confocal microscope. The results show that with transfection of CIB1 exogenous plasmid into 293T cells, more and more CIB1 were expressed,and there are obvious translocation of CIB1 from nuclear to cytoplasm. These results will highlight the understanding of CIB1 subculluar localization changes its potential roles. Key words:calcium binding protein; CIB1;293T cells; laser scanning confocal microscope