CAJ | 학술논문

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。
위건립은정표체증강형록색형광단백（EGFP）기인적산양태인성섬유세포（gFFC）주，채용조직괴첩벽법배양gFFC ，QIAGEN전염시제개도전염gFFC ，용G418제도법사선양성세포주。결과현시：배양적gFFC생장형태정상；24공배양판중세포회합도체70％～80％시，매60μL전염액내함DNA량0.4μg화 QIAGEN전염시제3μL ，매공내가입40μL전염액시능획득최가전염효과，재G418사선1개월후획득항성양성세포주。표명 EGFP기인재 gFFC중성공표체。
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 har-vested by tissue explants culture method and then transfected with the EG FP gene using the QIAGEN transfection reagent .The positive cells 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 40μL transfection medium ,which contains 0 .27μg DNA and 2 μl QIAGEN transfec-tion 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 .