中国生物工程杂志

The expression vector pGg-gfp containing gpd-Gf (615bp) promoter fragment from Grifola frondosa was constructed for detecting its functional activity by the fusion of promoter fragment to the reporter gfp 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 gfp 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.