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表达绿色荧光蛋白的猪胚胎生殖细胞分离

DOI: 10.7685/j.issn.1000-2030.2004.04.020, PP. 89-92

Keywords: ,胚胎生殖细胞,增强型绿色荧光蛋白,转染

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Abstract:

以质粒DNA为增强型绿色荧光蛋白(EGFP)的载体,对处于快速增殖期的8株猪胚胎生殖(EG)细胞(4~9代)进行转基因试验。用1μgDNA和2μLLipofecTamine2000转染EG细胞后,共获得4株转染EGFP基因的EG细胞系(EGEGFP),可发出强绿色荧光;EGEGFP细胞连续培养2周以上可分化为多种细胞类型。显微注射EGEGFP细胞至卵裂期胚胎、桑椹胚卵周隙内或囊胚腔中,共注射135枚胚胎,其中112枚存活并发育至囊胚(830%);86枚嵌合体胚培养后含有荧光细胞,其中57枚囊胚在内细胞团(ICM)上发现有EGEGFP细胞。

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