表达GFP/mCherry白色念珠菌的构建及其在与巨噬细胞互作中的应用

A Candida albicans strain expressing GFP/mCherry: construction and application in the research on macrophage- C.albicans interaction

摘要:

白色念珠菌(Candida albicans)被巨噬细胞吞噬的效率与被吞噬后的形态观察是研究白色念珠菌与巨噬细胞互作的重要内容。【目的】以野生型菌株SC5314为母本，构建能够表达绿色荧光蛋白(green fluorescent protein, GFP)/mCherry的白色念珠菌，应用于巨噬细胞与白色念珠菌互作的研究。【方法】通过生长与形态观察、细胞活性检测及小鼠系统性感染模型确定荧光蛋白的表达对菌株生长、形态与毒力的影响；在共培养条件下，通过流式细胞术及荧光显微镜检测巨噬细胞的吞噬率及白色念珠菌的形态变化。【结果】构建的菌株在表型上与野生型菌株一致，并可用于在共培养下测定巨噬细胞吞噬率的流式细胞术以及观察白色念珠菌的形态变化。【结论】表达荧光蛋白的菌株为研究巨噬细胞与白色念珠菌的互作提供了新方法。

Abstract:

The phagocytosis rate of macrophages and the morphological changes of Candida albicans are the key points in the research on the host-pathogen interaction. [Objective] To construct C. albicans strains expressing the green fluorescent protein (GFP) or mCherry from the wild type SC5314, and use them to study the interaction between macrophages and C. albicans. [Methods] The effects of fluorescent protein expression on the growth, morphology, and virulence of the strains in mice were determined. Under the co-culture condition, flow cytometry and fluorescence microscopy were employed to determine the phagocytosis rate of macrophages and the morphological changes of C. albicans, respectively. [Results] We constructed C. albicans strains expressing GFP or mCherry. These engineered strains had consistent phenotype with the wild type and could be used for flow cytometry to determine the phagocytosis rate of macrophages and the morphological changes of C. albicans in co-culture. [Conclusion] The constructed strains expressing fluorescent proteins provided a new method for studying the interaction between macrophages and C. albicans.