中文摘要：

通过对绿色荧光蛋白（GFP）布鲁氏菌弱菌株M5（GFP-M5）和S19（GFP-S19）侵染小鼠巨噬细胞（RAW264.7）及对其与胞内溶酶体、内质网、高尔基体初次结合所用时间进行测定,探讨分析两种布鲁氏菌弱毒株侵染小鼠巨噬细胞过程的荧光表征。将GFP-M5和GFP-S19分不同时间段分别侵染RAW264.7,利用激光共聚焦和流式细胞仪观察和检测。结果显示,GFP-M5和GFP-S19均构建成功。布鲁氏菌M5和S19及GFP-M5和GFP-S19侵染RAW264.7后胞内生存能力无明显差异。GFP-M5和GFP-S19侵染30min后均已进入小鼠巨噬细胞,2h分别到达溶酶体、内质网和高尔基体。而两种弱毒株在1、2、3、4h与各细胞器结合率相近。流式细胞仪检测结果显示,GFP-M5和GFP-S19侵染RAW264.7的GFP＋细胞含量无显著差异（P〉0.05）。结果提示,两种弱毒株在侵染进入宿主细胞的初期及侵袭能力并没有明显差异。

英文摘要：

The experiment was conducted to discuss the difference of binding time of green fluorescent protein B.melitensis M5（GFP-M5）and B.abortus S19（GFP-S19）infecting the mouse macrophagocyte（RAW264.7）,lysosome,endoplasmic reticulum and golgi body in the initial stage and compare the binding rate of GFP-M5,GFP-S19 with organelle in different timeline,respectively,by confocal laser scanning microscope（CLSM）and flow cytometry.The result showed that GFP-M5 and GFP-S19 were successfully constructed.The intracellular survival ability of Brucella M5,Brucella S19,GFP-M5 and GFP-S19 were not obvisouly affected after infecting RAW264.7.GFP-M5 and GFP-S19 could enter the macrophagocyte in 30 mins,and in 2h the Brucellacould reach lysosome,endoplasmic reticulum and golgi body.In addition,the binding time for two attenuated vaccine did not show differences in 1,2,3and 4h.The content of GFP＋cell produced by RAW264.7infected by GFP-M5 and GFP-S19 did not show significant differences（P〈0.05）.Therefore,the two strains did not have significant differences in the invasion ability in the initial stage of infecting host cell.