hEGF转基因表皮细胞的制备及其EGF蛋白表达  

The expression level of the transfected hEGF gene to cultured human epidermal cells

作　　者：董肇杨[1] 陈玉林[1] 戴方平[1] 郇京宁[1] 张素贞[1] 应康[1] 吕洛[1] 范清源[1] 方之扬[1] 

机构地区：[1]第二军医大学长海医院烧伤科,上海200433

出　　处：《中华整形外科杂志》2000年第3期177-179,共3页Chinese Journal of Plastic Surgery

基　　金：国家自然科学基金资助 !(批准号 3 9580 0 2 3 )

摘　　要：目的　应用G41 8筛选转染EGF基因后的转基因上皮细胞 ,并测定外源性基因在转基因细胞中的稳定表达。方法　选用构建的pBK Signal EGF质粒 ,在脂质体LipofectAMINE的介导下 ,转染培养 60 %～ 70 %连片的人表皮细胞 ,然后用G41 8进行筛选出稳定表达的细胞克隆。大量培养传代后 ,通过ELISA试剂盒检测细胞培养上清中表皮细胞生长因子 (EGF)外源性基因稳定表达水平。结果　①人表皮细胞生长因子hEGF质粒可成功转到人表皮细胞中 ;②通过G41 8可以筛选出含EGF的转基因细胞 ;③hEGF表达水平在细胞培养上清中可以测出。结论　①通过G41 8可以筛选出转EGF基因的表皮细胞 ;②采用体外培养的第Objective To study the stable expression levels of cultured human epidermal cells transfected with a hEGF plasmid.Methods The 60%～70% confluent cultured human epidermal cells were transfected with a pBK Signal EGF plasmid construct mediated by lipofectAMINE. After screening of G418 successfully transfected ,clone forming cells were picked and subcultured for several weeks. To detect hEGF expression levels and to determine long term transgenic stability a specific ELISA was used.Results (1) hEGF plasmid constructs were successfully introduced into cultured human epidermal cells. (2) Transgenic expression still persisted after four subcultures. (3) hEGF expression levels could be detected in the medium over a six week period.Conclusions The stable expression of hEGF was obtained after gene transfected with the pBK Signal EGF plasmid mediated by lipofectAMINE.

关 键 词：基因转梁 表皮生长因子 质粒 基因表达 

分 类 号：R965[医药卫生—药理学]