GFP-hDaxx融合蛋白在巨噬细胞中的表达与定位

The Expression and Localization of GFP-hDaxx Fusion Protein in Macrophages

目的 构建GFP -hDaxx融合蛋白真核表达载体pEGFP/hDaxx并在巨噬细胞中表达 ,为研究hDaxx对巨噬细胞凋亡及分泌活性的影响提供实验基础。 方法 将hDaxxcDNA片段亚克隆入载体pEGFP ,构建GFP -hDaxx融合蛋白真核表达载体。将质粒pEGFP/hDaxx转染巨噬细胞 ,Westernblotting鉴定表达产物 ,荧光显微镜观察GFP -hDaxx融合蛋白在细胞内的表达与定位。 结果 成功构建了GFP -hDaxx融合蛋白真核表达载体pEGFP/hDaxx ,GFP -hDaxx融合蛋白在巨噬细胞中表达并定位于细胞核。 结论 pEGFP/hDaxx在巨噬细胞中即时高表达GFP

Objective To construct eukaryotic expression vector pEGFP/hDaxx, which can express GFP-hDaxx fusion protein, and to study the effect that hDaxx affects macrophages apoptosis and its secretory activities. Methods hDaxx was subcloned into pEGFP vector and positive colony was selected. pEGFP/hDaxx was transfected into macrophages. Expression product of GFP-hDaxx fusion protein was analyzed by Western-blotting and observed localization under fluoscent microscope. Result pEGFP/hDaxx, which is an eukaryotic expression vector, was constructed successfully and GFP-hDaxx fusion protein were expressed ideally in macrophages and localized in nuclei. Conclusion GFP-hDaxx fusion protein immediately expressed in macrophages successfully and localized in nuclei.