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【目的】为能实时直观了解嗜肺军团菌感染细胞的过程,研究细菌在细胞内的变化及其与宿主细胞间的相互作用关系。【方法】通过基因敲除、克隆回补等重组构建绿色荧光蛋白(GFP)稳定高表达的嗜肺军团菌株,利用该菌株建立小鼠巨噬细胞Raw264.7的感染模型。【结果】通过荧光显微镜可实时观察细菌感染细胞的全过程,包括细菌在细胞内的形态变化、增殖和裂解宿主细胞等。【结论】重组菌可替代野生菌株在细胞感染中应用,为直观研究嗜肺军团菌与被感染细胞之间的相互作用关系,以及进行相关药物模型的制备、药物筛选、耐药机制研究等提供了新的手段。
【Objective】 To understand the process of Legionella pneumophila infection in real time and to study the changes of bacteria in the cells and their interactions with host cells. 【Method】 The Legionella pneumophila stable and highly expressed green fluorescent protein (GFP) was constructed by gene knockout, cloning and complement recombination, and the infection model of mouse RAW264.7 cells was established by using this strain. 【Result】 Fluorescence microscopy was used to observe the whole process of bacterial infection in real time, including the morphological changes of bacteria in cells, proliferation and lysis of host cells. 【Conclusion】 Recombinant bacteria could substitute wild strains for cell infection. To provide a theoretical basis for studying the interaction between Legionella pneumophila and infected cells, as well as preparation of related drug models, drug screening and drug resistance mechanisms A new means.