白色假丝酵母菌pACT1-GFP质粒的构建及表达

The construction and expression of Saccharomyces albicans pACT1-GFP plasmids

目的构建能够稳定表达绿色荧光蛋白(GFP)的白色假丝酵母菌菌株,以便对目的基因进行示踪。方法构建pACT1-GFP质粒,以白色假丝酵母菌CAI4菌株作为感受态进行转化培养后,分别观察绿色荧光蛋白在两相型下的表达情况。结果含有pACT1-GFP的白色假丝酵母菌菌株,99%在两相型下均有较高水平的荧光蛋白表达,且荧光强度没有明显差别。结论pACT1-GFP能够在白色假丝酵母菌体内稳定表达。

Objective To construct strains containing green fluorescent protein（GFP） to study gene regulation in Saccharomyces albicans cells during the infection process.Methods pACT1-GFP was constructed,and Saccharomyces albicans CAI4 was transformed.The expression of GFP in yeast and hyphal compartments was observed with micros-copy.Results 99% of Saccharomyces albicans cells containing pACT1-GFP fusion displayed significant fluorescence levels both in the yeast and hyphal compartments.The fluorescence intensity in two compartments had no obvious dif-ference.Conclusion pACT1-GFP can be expressed stably in the yeast cells.