调控子fur影响霍乱弧菌生物膜的形成(英文)  

The Role of Fur Involved in Bioflim Formation of Vibrio Cholerae

作　　者：陈秀琴[1] 蔡红艳[2] 张卫[1] 闫梅英[3] 高鹤[3] 段国贤[2] 闫志勇[1] 

机构地区：[1]青岛大学医学院,山东青岛266071 [2]河北联合大学基础医学院,河北唐山063000 [3]中国疾病预防控制中心传染病预防控制所,传染病预防控制国家重点实验室,北京102206

出　　处：《现代生物医学进展》2013年第15期2841-2845,共5页Progress in Modern Biomedicine

基　　金：Natural Science Foundation of China(30830008)~~

摘　　要：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： The ferric uptake regulator（Fur） regulates as both a repressor and an activator of numerous genes in Vibrio cholerae in response to maintain iron homeostasis.It has also been demonstrated that Fur plays an additional role in virulence in V.cholerae.Both biofilm formation and motility contribute to the virulence of V.cholerae.In this study we characterize Fur-dependent phenotypes in biofilm formation to forecast Fur-dependent transcription of virulence genes of V.cholerae.The expression of Fur protein make preparations for the further experiments,such as purified Fur protein,electrophoretic mobility shift assay（EMSA）,DNase I footprinting,RT-qPCR.Methods： The fur null mutant strain and the complemented mutant were constructed from the wild-type V.cholerae.Construction of the protein expression strain and preparation for purification His-tagged Fur（His-Fur） protein.The motility and biofilm formation was characterized for the fur null mutant strain and the complemented mutant.Results： The fur null mutant strain and the complemented mutant from V.cholerae were successfully constructed.The fur deletion leads to an enhance in biofilm formation and yet a decrease in mobility.Conclusion： This study predicts that Fur may affect the expression of numerous genes involved in flagellum biogenesis,extracellular polysaccharide and outer membrane protein synthesis.The phenotypic assays indicated that the transcriptional regulator Fur inhibited the V.cholerae biofilm production in vitro.Here we show that the transcriptional factor of Fur directly or indirectly regulatd the expression of some certain genes involved in the cellular processes of spread,infection,and colonization of V.cholerae and laid the foundation for further research.

关 键 词：霍乱弧菌 FUR His-Fur 生物膜形成 

分 类 号：Q735[生物学—分子生物学] R378.3[医药卫生—病原生物学]