中文摘要：

利用从灰树花菌丝体中克隆的gpd-Gf（615bp）启动子片段串联于报告基因gfp上游,构建启动子功能活性检测表达质粒pGg-gfp。采用PEG介导法把表达质粒pGg-gfp与辅助质粒pCc1001（含有trp1基因）共转化进色氨酸营养缺陷型的灰盖鬼伞粉孢子的原生质体中。经过选择培养基筛选、假定转化子的分子鉴定以及GFP荧光检测,结果表明：灰树花gpd-Gf启动子在灰盖鬼伞菌丝中具有较强驱动gfp基因表达的活性,在荧光显微镜和共聚焦显微镜下可以观察到转化子菌丝发出的强烈荧光。

英文摘要：

The expression vector pGg-gfp containing gpd-Gf （615bp） promoter fragment from Grifola was constructed for detecting its functional activity by the fusion of promoter fragment to the reporter gene. Co-transformation of plasmid pGg-gfp and plasmid pCc1001 which harbors the complementary gene trp1 was conducted by the PEG-mediated protoplast transformation of the oidia of LT2, a tryptophan auxotrophic strain of Coprinus cinereus. The putative trp ＋ transformants were obtained from the selective regeneration medium and confirmed by PCR detection, Southern blotting and green fluorescence analysis. The results indicated that gene was integrated into the gemone of LT2 strain. And the gfp-Gf promoter could drive the gfp gene expression in Coprinus cinereus. Strong green fluorescence acitivity was observed in the mycilia of Coprinus cinereus transformants under the fluorescent electronic microscope and laser scanning confocal microscope.