不同剪接形式鼠era在细胞内的定位

Intra-cellular Localization of Two Splicing Mouse era

为分析两种剪接形式鼠era在细胞内的定位,构建了era-EGFP融合表达载体pSMEGFP-meraW和pSMEGFP-meraS,然后用其转染鼠细胞系NIH3T3。荧光显微镜观察发现,两种剪接形式鼠era-EGFP融合蛋白均定位于细胞浆中的核周围。同时,应用免疫荧光细胞化学法分析了自然情况下,鼠细胞系野生型era蛋白在细胞内的分布,结果显示细胞核与细胞浆中均有分布。推测鼠era蛋白在细胞浆中合成修饰后,才转移至细胞核发挥作用。由于era-EGFP融合蛋白难以进入核内或需要较长时间,所以导致era-EGFP与野生型鼠era在细胞内定位不尽相同。

To study the intra-cellular localization of mouse era,era-EGF Pfusion expressive vectors is constructed： pSMEGFP -meraW and pSMEGFP-meraS, and then them is transfected into mouse fibroblast cell line NIH3T3. Two splicing mouse era-EGF Pproteins localized mainly in cytoplasm around cell nucleus is revealed by fluorescence microscopy analysis. The intra-cellular localization of mouse era in nature was analyzed by immunocytochemistry techniques and the results showed that mouse era localized in all subcellular parts. The reason of different results may be that era-EGFP fusion protein is difficult into cell nucleus.