关键词: 纤维素酶基因, 福寿螺, 毛柄金钱菌, gpd启动子

Abstract: In order to investigate the expression of multifunctional cellulase gene of Ampullarium crossean (mfc) in large filamentous fungi driven by glyceradehyde-3-phosphate dehydrogenase (gpd) promoter (Fvl) of Flammulina velutipes, a new eukaryotic expression vector containing F. velutipes gpd promoter (Fvl) and A. crossean cellulase gene (mfc) was constructed. The expression vector was inserted into the chromosome of tryptophan auxotrophic Corprinus cinereus by polyethylene glycol(PEG) mediation. The transformants were selected by PCR and Southern blotting. The transcription of mfc was confirmed by RT-PCR and the expression of mfc was further determined by measuring the activities of filter paper enzyme , carboxymethylcellulase and xylanase. The results showed that mfc driven by F. velutipes gpd promoter exhibited highly effective expression in engineered C. cinereus, and the strain with the highest cellulase activity was Cfvlm9, in which the filter paper enzyme, carboxymethylcellulase and xylanase activities were 21.5, 44 U/mL and 235 U/mL, respectively with 1.79, 1.6 and 2.97 fold increase as compared to the control strain.

Key words: cellulase gene, Ampullarium crossean, Flammulina velutipes, glyceraldehyde-3-phosphate dehydrogenase gene promoter