G418抗性HEK293细胞的培育

目的　培育具有G418抗性的HEK2 93细胞 ,用于建立猪内源性反转录病毒感染人HEK2 93细胞的模型。方法　通过脂质体转染的方法 ,将含有neo基因的质粒pIRESneo导入HEK2 93细胞中 ,利用G418的选择特性 ,对转染细胞进行压力筛选 ,并对其进行了PCR鉴定。结果　经 6 0 0 μg ml的G418压力筛选后 ,获得了抗性细胞克隆。抗性细胞的形态和生长速度与筛选前细胞没有差异 ,特异性核苷酸引物检测抗性细胞基因组DNA ,可以扩增出对应的核苷酸片段。结论　成功地培育了G418抗性HEK2 93细胞 ,为建立猪内源性反转录病毒感染人HEK2 93细胞的模型奠定了基础。

Establishment of a G418-Resistant HEK293 Cell Line

Objective In order to obtain a model of the HEK293 cells infected by porcine endogenous retroviruses PERV, by means of the G418|resistant HEK293 cell line established. Methods The plasmid pIRESneo containing neo gene was purified and transfected into HEK293 cells with Lipofectin. The transfected cells would be survived in the further culture medium containing G418 antibiotic as the neo gene could express a G418 resistant products. The identification of the G418 resistant HEK293 (G418 RHEK293) cells was conducted by polymerase chain reaction with the specific primers of neo gene. Results The G418 RHEK293 cell line was established successfully. There were no differences among G418 RHEK293 cells and normal HEK293 cells morphologically and propagatively. The neo gene DNA fragment could be amplified by PCR with specific primers in the genomic DNA of G418 RHEK293 cells. Conclusion The G418 RHEK293 cells were established with transfection of plasmid pIRESneo by Lipfectin, which may be useful as a cell model for the research of PERV.