Abstract:[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.