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目的:建立红色荧光蛋白mCherry在衣原体中稳定表达的方法，并使用荧光菌株Cm-mCherry构建有效的衣原体感染模型。方法:构建含有红色荧光蛋白mCherry的重组质粒pmCherry：：CM，利用氯化钙法将pmCherry：：CM转化至衣原体质粒缺失株CMUT中，经过筛选与纯化得到荧光菌株Cm-mCherry。Cm-mCherry感染HeLa229细胞后观察包涵体中红色荧光的发光情况，并与间接免疫荧光染色法比较，评估细胞感染模型构建是否成功；Cm-mCherry感染小鼠后，通过测定下生殖道载菌量、冷冻切片观察上生殖道衣原体红色荧光、分离生殖道评价输卵管积水发病情况，确定Cm-mCherry的感染力、侵袭力与致病性，评估动物感染模型构建是否成功。结果:红色荧光蛋白mCherry在转化株Cm-mCherry中稳定表达，红色荧光蛋白标记法与间接免疫荧光染色法一致度高；Cm-mCherry小鼠感染模型中，感染率为100%(10/10)，下生殖道载菌量可稳定保持高水平，在上生殖道冷冻切片中可观察到带有红色荧光的包涵体，输卵管积水发病程度与Cm野生株差异无统计学意义(n P>0.05)，证实Cm-mCherry有较好的感染力、侵袭力与致病性。n 结论:成功建立红色荧光蛋白mCherry在衣原体中稳定表达的方法，转化后的荧光菌株Cm-mCherry有较好的感染力、侵袭力与致病性，可用于衣原体细胞、动物感染模型的构建，为研究衣原体感染机制提供良好的实验工具。“,”Objective:To establish a method for stable expression of red fluorescent protein mCherry in n Chlamydia muridarum (Cm) and construct an effective n Chlamydia infection model using the fluorescent strain (Cm-mCherry).n Methods:A recombinant plasmid (pmCherry: : Cm) containing mCherry gene was constructed and transformed into CMUT, a plasmid-deficient n Chlamydia strain, using calcium chloride. The fluorescent strain Cm-mCherry was selected and purified. Red fluorescence in the inclusion bodies was observed after infecting HeLa229 cells with Cm-mCherry and the method of indirect immunofluorescence (IIF) was used as a comparison to evaluate the establishment of cell infection model. The infectivity, invasiveness, and pathogenicity of Cm-mCherry and the establishment of animal infection model were evaluated by measuring the number of n Chlamydia in the lower genital tract, observing the red fluorescence in the upper genital tract and analyzing the incidence of hydrosalpinx following infecting mice with Cm-mCherry.n Results:The red fluorescent protein mCherry was stably expressed in Cm-mCherry. Red fluorescent protein was highly consistent to IIF for labelling n Chlamydia. In the mice infection model, the infection rate was 100% (10/10). The load of n Chlamydia in the lower genital tract was stable and maintained at a high level. The inclusion bodies with red fluorescence were observed in the frozen sections of the upper genital tract. No significant difference in the severity of hydrosalpinx was found between Cm-mCherry and wild-type Cm infection groups (n P>0.05). These results confirmed the good infectivity, invasiveness and pathogenicity of Cm-mCherry.n Conclusions:Stable expression of the red fluorescent protein mCherry in n Chlamydia was achieved. The transformed strain Cm-mCherry had good infectivity, invasiveness and pathogenicity and could be used to construct the cell and animal models of n Chlamydia infection, which would be conducive to study the mechanism of n Chlamydia infection.n