【摘要】 采用绿色荧光蛋白 (GFP)标记技术研究钙离子钙调素依赖性蛋白激酶Ⅱ (CaMKⅡ )在HeLa细胞中的分布 .为了研究细胞内钙离子钙调素信号的下游作用物 ,我们首先构建了CaMKⅡ与GFP的融合基因 ,磷酸钙沉淀法将pCaMKⅡ -GFP重组载体导入HeLa细胞 .通过荧光显微镜观察 ,发现在细胞间期 ,CaMKⅡ -GFP融合蛋白主要分布于细胞核中 ,细胞质中亦有少量分布 ,这一分布不同于绿色荧光蛋白在间期细胞内的均匀分布 .同时讨论了与间接免疫荧光染色方法相比 ,GFP技术在功能蛋白定位研究上的应用优势 .更多还原

【Abstract】 The distribution of calcium/calmodulin dependent protein kinase Ⅱ (CaMKⅡ) in HeLa cell was studied by using green fluorescent protein (GFP) as a tag.In order to study the downstream components of calcium calmodulin signaling, we fused the gene of CaMKⅡ with that of GFP and we expressed it in HeLa cells. Observation with fluorescent microscope confirmed expression of the fusion protein. We found that, in interphase HeLa cells CaMKⅡ mainly concentrated in the nucleus and little in the cytoplasm,a pattern different from the equal distribution of GFP .The superiosity of this technique over indirect immunofluorescence staining was also discussed.更多还原