利用cryparin基因的启动子构建板栗疫病菌高效表达载体

Construction of Highly Efficient Over Expression Vector Containing cryparin Promoter for Cryphonectria parasitica

低毒病毒/板栗疫病菌是研究植物病原菌致病机理和病毒与宿主相互作用的一个优秀模式系统。本研究克隆了板栗疫病菌转录水平最高的cryparin基因的启动子,并构建了由该启动子控制的表达载体。构建的载体能成功表达GFP蛋白。利用该载体表达积累量较高的CHV1-Euro7病毒的病毒量控制基因,能提高细胞内CHV1-EP721的积累量,反式互补效率从常用的gpd启动子控制的低于10%提高至67%和80%。高效表达载体的成功构建,为研究板栗疫病菌功能基因以及低毒病毒与宿主板栗疫病菌的相互作用提供了新的工具。

Hypovirus/Cryphonectria parasitica is an excellent model system to study the pathogenic mechanism of fungus and the virus-host interaction. In this study, a highly efficient over-expression vector with the promoter from the most dominantly expressed gene cryparin was constructed. Green fluorescence protein （GFP） can be suc- cessfully expressed in the transformants, and an improved complementation rate of 67% and 80% from less 10% was achieved by expression of the determinant from the higher titer virus CHV1-Euro7 in the cell infected with the lower titer virus CHV 1-EP721 as compared with commonly used gpd promoter. The availability of the highly efficient over-expression vector provides a new tool to study the function of genes in Cryphonectria parasitica and virus-host interaction.