中文摘要：

目的构建新生隐球菌的PMT4基因缺陷株及新生隐球菌生物膜的体内、外模型；研究PMT4基因对生物膜形成的影响。方法采用PCR介导的长侧翼同源重组的方法敲除新生隐球菌H99的PMT4基因；采用基础培养基96孔板培养的方法建立生物膜体外模型；兔中心静脉插管、管内放置聚苯乙烯薄膜的方法建立生物膜动物模型；用倒置显微镜、共聚集激光扫描显微镜、MTT、CFU计数等方法研究PMT4缺陷株与野生株生物膜的异同。结果新生隐球菌在体内、外模型中均能形成生物膜；PMT4缺陷株与野生株生物膜在生物量和结构方面存在明显差异。结论本实验的生物膜动物模型可行；PMT4基因缺陷可造成隐球菌生物膜代谢活性下降，并形成假菌丝样结构。

英文摘要：

Objective To build a PMT4 mutant of C. neoformans H99 and a new animal model of C. neoformans biofilms. To investigate the difference between H99 and it＇s PMT4 mutant in formation of biofilms. Methods Long Flanking Homology （LFH-） PCR was used to obtain PMT4 mutant. Animal model was set up by setting a catheter （with polystyrene film in lumens） into rabbit central venous. Inverted phase contrast microscope, CLSM and CFU counting were applied in this study. Results Both H99 and PMT4 mutant were able to form biofilms in the animal and in vitro model. The biofilms of H99 and PMT4 mutant were similar, but the latter had less amount of cells and CFU than the former in the same phrase. Mycelioid structure appeared in biofilms of PMT4 mutant 24h after inoculation. Conclusions The data suggested that PMT4 mutant biofilms had less biomass and proliferative cells than wild strain. The absence of PMT4 gene may result in the change of biofilms＇structure.