Fraser D, Diatloff-Zito C, Moustacchi E
Institut Curie-Biologie, URA 1292 du CNRS, Paris, France.
Mutat Res. 1991 Jul;263(3):165-71. doi: 10.1016/0165-7992(91)90057-b.
An electroporation protocol for the successful transfection of mouse lymphoblastoid cells has been developed. Two cell lines, a normal and a mutant sensitive to DNA cross-linking agents, were used. The optimum conditions of electroporation in terms of uptake of the fluorescent dye lucifer yellow coupled with low toxicity were established. Subsequently, these conditions were used to achieve stable transfection by a plasmid expression vector. The plasmid integration patterns were determined by Southern blot analysis.
已开发出一种用于成功转染小鼠淋巴母细胞的电穿孔方案。使用了两种细胞系,一种是正常细胞系,另一种是对DNA交联剂敏感的突变细胞系。根据荧光染料路西法黄的摄取情况并结合低毒性确定了电穿孔的最佳条件。随后,利用这些条件通过质粒表达载体实现稳定转染。通过Southern印迹分析确定质粒整合模式。