带GFP标记嗜肺军团菌的构建和在细胞感染中的应用  

Construction and application of green fluorescent protein-tagged Legionella pneumophila in cell infection

作　　者：熊丽娜[1,2] 赵红波[2] 莫自耀[2] 石磊[1] 

机构地区：[1]华南理工大学,广东广州510641 [2]广州医科大学附属第一医院广州呼吸疾病研究所呼吸疾病国家重点实验室,广东广州510120

出　　处：《微生物学通报》2014年第11期2325-2331,共7页Microbiology China

基　　金：国家重点实验室自主选题基金项目

摘　　要：【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 study the infection by Legionella pneumophila and the relative biological activities between bacteria and host cells in real time.[Methods] We constructed a strain of L.pneumophila that can stably express green fluorescent protein by method of gene knockout and retro-complementation,and established an infection model with murine Raw264.7 macrophage.[Results] The recombinant strain was applied in cell infection successfully.The entire process of infection can be observed in real time by fluorescence microscope,including the variety of bacterial cell morphology,intracellular proliferation,and lysing the host cells.[Conclusion] Our result provides a new mean to study the relationship between L.pneumophila and infected cells,the preparation of some models related to the drugs,and the drug screening and the mechanism of drug resistance.

关 键 词：嗜肺军团菌 绿色荧光蛋白 感染 

分 类 号：R378[医药卫生—病原生物学]