【摘要】 目的研究脂质体转染方法将质粒DNA导入小鼠受精卵中的可行性。方法用免疫细胞化学方法在受精卵内检测pAkt(Ser473)蛋白的分布。用脂质体转染的方法将质粒FLAG-PIPP或FLAGvector导入受精卵,再用Western Blot检测两组卵细胞中pAkt(Ser473)的表达量。用荧光显微镜检测GFP-PH/ARNO质粒在卵细胞中的表达同时观察胰岛素对其影响。结果 pAkt(Ser473)抗体成功穿过透明带表达在受精卵内,细胞膜表达最强。与FLAGvector转染的受精卵相比,pAkt(Ser473)在转染FLAG-PIPP质粒的受精卵内表达量明显减少。与PIP3结合的GFP-PH/ARNO绿色荧光以细胞核表达为主,并且胰岛素可以使荧光增强。结论脂质体转染方法可成功地将质粒DNA穿过透明带导入小鼠受精卵内。更多还原

【Abstract】 Objective To explore the possibility that plasmids DNA is transfected directly into mouse fertilized eggs by lipofection in rat. Methods The distribution of pAkt Ser473 was observed with immunocytochemical analysis in mouse fertilized eggs. FLAG-PIPP or FLAG vector were transfected into mouse fertilized eggs and then Western Blot was used to detected their corresponding expression of pAkt (Ser473). GFP-PH/ARNO was transfected into eggs and observed by fluorescent microscope. Results The expression of pAkt Ser473 was observed distinctly in zona pellucida(ZP)-intact fertilized eggs, the expression level of pAkt Ser473 was decreased in the mouse transfected with FLAG-PIPP than in those eggs transfected with FLAG vector. The green fluorescence of GFP-PH/ARNO combined with PIP3 was also observed in the area of nuclei of eggs. Conclusion Lipofection is possible method for the delivery of plasmids DNA into mouse fertilized eggs.更多还原