担子菌基因启动子在构巢曲霉遗传转化中的应用OA

Promoters and terminators are key regulatory elements for expressing exogenous genes in genetic function studies.Due to significant genetic differences among fungal phyla,gene expression regulatory elements are generally used only within the same species or between highly similar species.Aspergillus nidulans serves as a model species for studying fungal genetic development and is also widely used in synthetic biology research for exogenous protein expression.Currently,genetic transformation in A.nidulans primarily utilizes promoters from its own constitutive gpd or tef genes,for protoplast transformation mediated by PEG.In this study,an Agrobacterium tumefaciens-mediated approach was employed to genetically transform A.nidulans apospores using the expression plasmid P020 originally developed for basidiomycetes.This plasmid contains a carboxin resistance gene expression cassette driven by the self-promoter derived from Pleurotus eryngii,and an eGFP expression cassette driven by the gpd promoter from Hypsizygus marmoreus.P.eryngii-derived carboxin gene driven by its own promoter efficiently screens positive transformants in A.nidulans via Agrobacterium tumefaciens-mediated transformation,while the gpd gene promoter from P.eryngii drives eGFP expression in A.nidulans mycelium.The above studies demonstrate that regulatory elements such as promoters and terminators from P.eryngii and H.marmoreus can successfully drive exogenous gene expression and exert protein function in A.nidulans.This study provides insights for utilizing gene regulatory elements from distantly related species in fungal expression systems.