脂质体载体法转染FPRL1基因到HEK293细胞及其鉴定

Transfection of FPRL1 Gene into HEK293 Cells by Liposome Method and Its Identification

[目的]验证脂质体载体法将FPRL1基因有效转染到HEK293细胞。[方法]G418筛选，RT-PCR和趋化试验。[结果]用脂质体载体法将FPRL1基因转染HEK293细胞，G418筛选，3周形成集落状细胞克隆；RT-PCR以FPRL1特异性引物扩增，凝胶电泳可见1条约700bp的目的片段，对照组为阴性；趋化实验显示HEK293/FPRL1细胞在一定浓度的fMLP的刺激下有趋化活性，而HEK293细胞无反应。[结论]初步证实脂质体载体法能有效地将FPRL1基因导入HEK293细胞，且FPRL1在细胞中有表达。

ObjectivesTo identify successful transfection of FPRL1 gene into HEK293 cells by liposome method. MethodsG418 filtration, RT PCR and chemotaxis test were used.ResultsWith liposome method, FPRL1 gene was transfected into HEK293 cells;then G418 filtration was performed. The colony like cell clones were formed 3 wks later. RT PCR demonstrated that FPRL1 gene was transfected successfully and expressed in HEK293/FPRL1 cells. A band of object fragment with about 700 bp in size was visible on gel electrophoresis, whereas negative result was obtained in control group. Chemotaxis suggested that FPRL1 was expressed on the surface of HEK293/FPRL1 cells, which displayed chemotactic response to a certain concentration of fMLP; while HEK293 cells were non responsive.ConclusionThe results of RT PCR and chemotaxis test confirm that successful transfection of FPRL1 gene into HEK293 cells with its intracellular expression is achieved by liposome method.