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目的:铁摄取调节蛋白Fur,其在细菌体内维持铁代谢稳态中担任着抑制剂和激活剂的双重角色,已有研究证实Fur还是霍乱弧菌的毒力调控子。研究证实生物膜的形成和游动性与霍乱弧菌的毒力相关,因此我们通过一系列表型实验分析Fur蛋白对霍乱弧菌生物膜形成影响,并预测依赖Fur与霍乱弧菌致病相关的基因,为后续研究做准备。方法:基于自杀质粒缺失霍乱弧菌的Fur基因,并构建相应回补株及蛋白表达株;通过表型实验,比较霍乱弧菌野生株和fur突变株在细菌运动能力、生物膜形成。结果:成功构建霍乱弧菌fur缺失株,回补株和蛋白表达株,His-Fur蛋白在上清液表达。表型实验表明fur突变株的菌株游动性降低,生物膜形成能力增强。结论:因此我们得出Fur可能与其它调控子一起,调控霍乱弧菌的表多糖,TCP和鞭毛蛋白的合成,从而抑制霍乱弧菌生物膜的合成。转录调控子Fur直接或间接调控一些与环境生存和致病相关的基因,对霍乱弧菌的传播、侵染、定殖等过程发挥作用奠定了基础。
OBJECTIVE: Iron uptake regulatory protein Fur, which plays a dual role as an inhibitor and activator in the maintenance of iron homeostasis in bacteria, has been shown to be virulence regulator of V. cholerae. Studies confirm biofilm formation and motility associated with Vibrio cholerae virulence, so we analyzed the effect of Fur protein on Vibrio cholerae biofilm formation through a series of phenotypic assays and predicted that the dependence of Fur on the pathogenesis of V. cholerae Of the genes for the follow-up study to prepare. Methods: The Fur gene of Vibrio cholerae was deleted based on the suicide plasmid, and the corresponding strains of complement and protein were constructed. Through the phenotypic experiments, the bacterial activity and biofilm formation were compared between the wild and the mut of Vibrio cholerae. Results: The Vibrio cholerae fur mutant strain, the complement strain and the protein expression strain were successfully constructed, and His-Fur protein was expressed in the supernatant. The phenotypic experiments showed that the fur mutant strains have lower mobility and enhanced biofilm formation ability. CONCLUSIONS: We therefore conclude that Fur, along with other regulators, regulates the synthesis of V. cholerae surface polysaccharides, TCP and flagellin, thereby inhibiting the synthesis of V. cholerae biofilms. Transcriptional regulator Fur directly or indirectly regulate some genes related to environmental survival and pathogenicity, and lay the foundation for the process of Vibrio cholerae dissemination, infection and colonization.