项目名称: 鼠疫耶尔森氏菌噬菌体Yep-phi受体结合蛋白与受体相互作用的分子机制研究
项目编号: No.31200137
项目类型: 青年科学基金项目
立项/批准年度: 2013
项目学科: 微生物学、植物学
项目作者: 赵向娜
作者单位: 中国人民解放军军事医学科学院
项目金额: 26万元
中文摘要: 鼠疫耶尔森氏菌噬菌体因对宿主的高度特异性和敏感性,对临床诊断具有重要意义。随着多重耐药性鼠疫菌的出现,噬菌体有可能成为用于预防和治疗鼠疫菌感染的抗生素替代品。噬菌体裂解宿主的特异性主要取决于噬菌体与细菌表面受体的识别与吸附。因此,揭示噬菌体与受体相互作用的分子机制是了解噬菌体裂解机制的重要基础。本研究拟以我国鼠疫菌诊断用噬菌体Yep-phi为对象,采用亲和层析、表面等离子共振技术、截短体构建和定点突变等手段,鉴定噬菌体受体结合蛋白与受体,评价鼠疫菌受体突变株与回补株的裂解效率,进而验证和分析噬菌体受体结合蛋白和受体的相互作用和作用的结构域。最终阐述噬菌体与受体结合的分子机制,为鼠疫的噬菌体防治提供理论依据。
中文关键词: 鼠疫耶尔森氏菌;噬菌体;受体结合蛋白;受体;
英文摘要: Yersinia pestis phages play an important role in clinical diagnosis due to their high specificity and sensitivity to its host. With the emergence of multiple drug-resistant Yersinia pestis, phages could be used as alternatives to antibiotics for the prevention and treatment of plague. Phage's host specificity depends largely on the recognition and adsorption of the phage to its host surface receptors. Therefore, it is critical imporatnt to understand the process of phage lysis exploring the mechanism of phage receptor-binding protein and receptors interaction. In this study, Yersinia pestis diagnosis phage Yep-phi, isolated in China, will be studied to identify the phage receptor-binding protein and its receptors and evaluate the plaquing efficiencies of phage on Yersinia pestis receptor knockout mutants and trans-complemented clones, and the necessary domains of the interaction will be identified and validated using affinity chromatography, surface plasmon resonance technology, construction of truncated receptor binding protein and receptors, site-directed mutagenesis and so on. Finally, the mechanism of phage adsorption to its receptors will be determined. This study will provide a theoretical basis for plague prevention and control.
英文关键词: Yersinia pestis;phage;receptor-binding protein;receptor;