CAJ | 학술논문

目的：应用Nucleofector II核转染仪将外源基因EGFP导入西藏小型猪胚胎成纤维细胞（PEFs），并与脂质体转染的方法进行比较，寻求一种高效转染猪胚胎成纤维细胞的方法。方法使用 Lonza 公司的Nucleofector II核转染仪，参考已报道的转染程序与相应的转染试剂，将绿色荧光蛋白（ GFP）基因导入西藏小型猪PEFs，并与脂质体转染的方法进行对比研究，比较两种方法的转染效率。结果转染5 h后，核转染仪组镜下即可见绿色荧光，转染效率高达80％，远远高于脂质体转染的方法。结论用Nucleofector II核转染仪能实现西藏小型猪PEFs的高效转染，为高效制备转基因克隆西藏小型猪提供了可靠的方法。
목적：응용Nucleofector II핵전염의장외원기인EGFP도입서장소형저배태성섬유세포（PEFs），병여지질체전염적방법진행비교，심구일충고효전염저배태성섬유세포적방법。방법사용 Lonza 공사적Nucleofector II핵전염의，삼고이보도적전염정서여상응적전염시제，장록색형광단백（ GFP）기인도입서장소형저PEFs，병여지질체전염적방법진행대비연구，비교량충방법적전염효솔。결과전염5 h후，핵전염의조경하즉가견록색형광，전염효솔고체80％，원원고우지질체전염적방법。결론용Nucleofector II핵전염의능실현서장소형저PEFs적고효전염，위고효제비전기인극륭서장소형저제공료가고적방법。
Objective To transfect EGFP gene to porcine embryonic fibroblasts ( PEFs) of Tibetan miniature pigs by Lonza Nucleofector II machine and compare the tansfection efficiency between this method and the lipofection method. Method A plasmid carrying green fluorescent protein ( GFP) was transfected into PEFs of Tibetan miniature pigs via the Lonza Nucleofector II machine ( program U020) and by Lipofectamine 2000.Results 5 hours after nucleofection, green fluorescence was observed, indicating 80%transfecting efficiency in the nucleofection group, which is significantly higher than the lipofection group. Conclusion Nucleofector II machine can efficiently transfect PEFs, provides a reliable method for efficiently generate transgenic Tibetan minipigs.