绿色荧光蛋白在鸡胚成纤维细胞中的表达

应用阳离子脂质体介导法,将含绿色荧光蛋白(GFP)基因的质粒pEGFP-C1转染到培养至第二代的鸡胚成纤维细胞(CEF)中,同时使用活体DNA染料Hoechst3334(2H342)对CEF细胞核荧光染色,通过荧光倒置显微镜和RT-PCR方法检测GFP的表达产物。在荧光倒置显微镜下可见CEF的胞质和核均呈现绿色荧光,且细胞核的绿色荧光强度强于细胞质,在H342作用下,细胞核呈现蓝色荧光;转染细胞中转录产物经RT-PCR扩增后,在凝胶上可见与GFP基因片段分子量大小一致的条带。可见,GFP基因可以被转染至CEF中,并在CEF中表达。

Expression of Green Fluorescent Protein in Chicken Embryo Fibroblast

The plasmid pEGFP-C1 containing green fluorescent protein(GFP)gene was transfected into chicken embryo fibroblast(CEF)of the second generation using lipofectin method. The nucleolus was stained by DNA dye Hoechst 33342(H342). The expression product of GFP gene was observed under fluorescence microscope and detected by RT-PCR. Results were that CEF expressed the product of green fluorescence,and the green fluorescence of nucleolus was more intensive than that of cytoplasm. The nucleolus expressed blue fluorescence by H342 staining. The expression of GFP was detected by RT-PCR and the fragment was corresponded with that predicted Mr of GFP. It suggested that GFP gene can be transfected into the CEF and expressed effectively in the CEF.