人肝再生增强因子和荧光蛋白基因在绵羊成纤维细胞中的表达

ALR and EGFP gene expression in sheep fetal fibroblasts cell

本实验利用LipofectAMINETM介导pEGFP/ALR载体转染sFFCs,经G418筛选,10d后共形成38个单克隆细胞,细胞用于提取蛋白进行聚丙烯酰胺凝胶电泳及免疫组化检测。结果表明:3个荧光单克隆细胞,聚丙烯酰胺凝胶电泳可见与预期相符的57KD大小的融合蛋白,免疫组化检测可见ALR蛋白分布于sFFCs胞质和胞核,且细胞核含量多于细胞质。

Sheep fetal fibroblasts were transfected with pEGFP-C1/ALR using LipofeetAMINETM.The transfected cells were incubated in the selected media containing 800ug/ml G418.Totally 38 clones were formed 10 days later. SDS-PAGE results indicated that there was specific 57KD protein band in the positive clons and a negative clon. Immunohistochemical analysis also indicated the expression of ALR gene were distributed in cytoplasm and nuclear, which matches the expression of GFP as well.Therefore,GFP can be used as a marker for target gene in the screening of transgenic cells and tissues.