Abstract:

Sheep fetal fibrobast cell were transfected with pEGFP-C1 by using Lipofectamine^TM or Fugene-6 substance respectively. The impacts of DNA concentration, concentration of transfeetion agents, exposure time and incubation time on transfection efficiency were carefully compared. The transfected cells were incubated in DMEM/F12 media containing G418. The results showed that the efficiency of transgenic with LipofectamineTM were excelled Fugene-6. Chromosome analysis shows no remarkable difference between transgenic and normal cells. LipofectamineTM transfection method developed in this paper can be easily adopted by most research labs and will provide a basic reference for transfection of sheep fetal flbroblast ceils with other gene and transgenie nuclear transfer.

Key words: sheep fetal fibroblast cell , liposome , gene transfer, chromosome

摘要：

分别用Lipofectamine^TM和Fegene-6介导质粒pEGFP-C1转染体外培养的绵羊胎儿成纤维细胞（sheepfetal fibroblast cells,sFFCs）,比较了DNA浓度、转染试剂用量以及细胞暴露于DNA、转染试剂中作用时间对转染效率的影响,通过含G418的DMEM/F12培养液筛选得到转基因单克隆细胞。结果表明脂质体转染试剂Lipofectamine^TM转染效率优于Fegene-6。另外,对转基因细胞进行了染色体核型分析。结果表明,转基因细胞中二倍体核型占74.5%,与对照组比较没有显著性差异。以上研究为其他基因转染sFFCs以及利用体细胞克隆法生产转基因绵羊提供了参考依据。

关键词: 绵羊胎儿成纤维细胞 脂质体 转基因 染色体