【摘要】 将质粒GH12596中的果蝇SNF1AcDNA用PCR方法扩增,并与pEGFP N2质粒中的荧光蛋白GFP编码序列连接入果蝇转基因载体pUAST上,构建了UAS SNF1A GFP质粒.转入果蝇细胞内,利用mini white标记基因筛选,得到3个独立的转基因果蝇品系.将这些转基因果蝇品系分别定位平衡,并用单果蝇PCR方法加以佐证.然后将转基因品系分别与eyGAL4,actin GAL4,pGMR GAL4等3个GAL4果蝇品系交配,在荧光显微镜下检测到该基因在果蝇中的表达.从而确认了用显微注射技术能够得到稳定的转基因果蝇模型,为开展用果蝇GAL4 UAS系统大规模研究基因功能奠定了基础.更多还原

【Abstract】 The recombinant vector UASSNF1AGFP harboring a fusion gene of Drosophila gene SNF1A and GFP was constructed. The construct was microinjected into the eggs of Drosophila and three independent transgenic fly lines were obtained. After balancing these fly lines, the integration of the transgene into Drosophila genome was further confirmed by PCR. The transgenic fly lines were then crossed with three different GAL4 fly lines respectively. Fluorescence analysis detected the expression of the transgene. The results show that foreign gene transgenic fly lines have been successfully constructed.更多还原