关键词: 油菜子饼肥, 油菜子饼肥, 土壤酶活性, 烤烟, 质量

Abstract: Yeast-like conidia of Tremella fuciformis were used as the material. Plasmid pEGFP and pCAMBIA1300 were digested by Hind III and EcoRⅠ restriction enzymes and ligated to transform. We constructed Tremella fuciformis expressed vector pCB-BEGFP contained enhanced green fluorescent protein gene which was regulated by gpd promoter of Agaricus bisporus. Electroporation was performed to transfer plasmid DNA of pCB-BEGFP into yeast-like conidia from Tremella fuciformis. The chromosomal DNAs of the four transformants and the nontransgenic strain as the control were used as the templates for the PCR using the EGFP-specific primers. When the products were analyzed by the agarose gel electrophoresis, the expected 750bp amplified bands appeared. By fluorescence microscopy, distinct green fluorescence could be observed from the colonies of YLCs grown on RM agar plates and could not be observed from the colonies of YLCs which did not contain plasmid pCB-BEGFP. Expression of EGFP was detected by SDS-PAGE analysis of TCP of one transformants, as expected, a dark band of the protein was visualized at 27kD by Coomassie Brilliant Blue staining. These results indicated the EGFP gene had been integrated into the genome of transgenic Tremella fuciformis strains and was expressed successfully by regulation of gpd promoter of Agaricus bisporus.