绿色荧光蛋白基因在山羊胎儿成纤维细胞中的表达  

Expression of EGFP Gene in Goat Fetal Fibroblast Cells

作　　者：楚秋霞[1] 王二耀[1] 吴姣[2] 陈付英[1] 王治方[1] 辛晓玲[1] 施巧婷[1] 冯亚杰[1] 李文军[1] 魏成斌[1] 徐照学[1] 

机构地区：[1]河南省农业科学院畜牧兽医研究所,河南郑州450002 [2]河南农业大学牧医工程学院,河南郑州450002

出　　处：《河南农业科学》2013年第10期142-145,共4页Journal of Henan Agricultural Sciences

基　　金：河南省农业科学院基础研究专项资金项目;国家转基因肉羊新品种培育专项(2013ZX08008003-002)

摘　　要：为建立稳定表达增强型绿色荧光蛋白(EGFP)基因的山羊胎儿成纤维细胞(gFFC)株,采用组织块贴壁法培养gFFC,QIAGEN转染试剂介导转染gFFC,用G418梯度法筛选阳性细胞株。结果显示:培养的gFFC生长形态正常;24孔培养板中细胞汇合度达70%～80%时,每60μL转染液内含DNA量0.4μg和QIAGEN转染试剂3μL,每孔内加入40μL转染液时能获得最佳转染效果,在G418筛选1个月后获得抗性阳性细胞株。表明EGFP基因在gFFC中成功表达。In order to establish the goat fetal fibroblasts cell （gFFC） clones stably expressing the enhanced green fluorescent protein （EGFP） gene,the goat fetal fibroblast cells （gFFC） were harvested by tissue explants culture method and then transfected with the EGFP gene using the QIAGEN transfection reagent. The positive ceils with transgenes were screened using different concen- trations of G418. The results showed that the propagation of gFFC was normal and the highest transfec- tion efficiency was achieved when using the 70 ^--80 ~ confluent cells in the 24-well plate cultured with supplement of 40 μL transfection medium,which contains 0.27 μg DNA and 2 μl QIAGEN transfection reagent. One month later,the drug-resistant cell clones gradually formed under the selection of G418. The results indicated that the EGFP was expressed efficiently in gFFC

关 键 词：山羊胎儿成纤维细胞 转染 EGFP基因 表达 

分 类 号：Q813.11[生物学—生物工程]