GFP-hDaxx融合蛋白在真核细胞中的表达与鉴定

目的：构建GFP-hDaxx融合蛋白真核表达载体pEGFP/hDAXX，并在HeLa细胞中表达，为进一步研究hDaxx在细胞凋亡、病毒感染中的作用提供实验基础。方法:用T4DNA连接酶将hDaxx亚克隆入载体pEGFP，筛选阳性克隆。将质粒pEGFP／hDaxx转染HeLa细胞，Western-Blotting鉴定表达产物，荧光显微镜观察GFP-hDaxx融合蛋白在细胞内的定位。结果：成功构建了GFP-hDaxx融合蛋白真核表达载体pEGFP／hDaxx，GFP-hDaxx融合蛋白成功表达并定位HeLa细胞核。结论:GFP-hDaxx融合蛋白成功地在真核细胞表达并定位于细胞核。

Expression and Identification of GFP- hDaxx Fusion Protein in Eukaryotic Cells

Objective To construct eukaryocytic expression vector pEGFP/hDaxx, which can express GFP -hDaxx fusion protein. Methods Subclone hDaxx into pEGFP vector and select positive clone. HeLa cells were transfected with eukaryotic expression vectors encoding GFP and GFP - hDaxx fusion protein. GFP - hDaxx fusion protein was analyzed by Western - blotting and fluoscent localization in HeLa cells. Results pEGFP/hDaxx, which is an eukaryocytic expression vector, was successfully constructed and GFP - hDaxx fusion protein was ideally expressed in HeLa cells, what is more, GFP - hDaxx fusion protein was observed in nuclei under fluoscent microscope . Conclusion GFP - hDaxx fusion protein expressed successfully in eukaryotic cells and localized in nuclei.