溶藻弧菌菌落相变关键小RNA的鉴定和调控机制研究

项目名称： 溶藻弧菌菌落相变关键小RNA的鉴定和调控机制研究

项目编号： No.31272697

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 陈偿

作者单位： 中国科学院南海海洋研究所

项目金额： 78万元

中文摘要： 溶藻弧菌是南方海域养殖动物弧菌病的主要病原之一，前期研究中我们发现菌落相变是决定其环境和宿主适应性的重要机制，不同菌落形态的菌株间生物膜形成能力、鞭毛合成与运动能力、碳源和氮源利用效率、对抗生素和胁迫条件反应能力等呈现显著差异，遗传分析结果表明菌落相变涉及复杂的信号传导和基因调控网络，其中非编码RNA（ncRNA）分子伴侣hfq及相关小RNA可能是控制此现象的决定因子,hfq基因缺失突变后改变了菌落形态，导致致病力消失，影响生物膜形成、物质转运与利用、抗胁迫能力等关键生理生化途径，显示小RNA对溶藻弧菌基本生命过程，尤其是对环境适应性和致病力等方面的重要调控作用。本项目拟在此基础上，通过转录组测序和生物信息分析鉴定溶藻弧菌小RNA及其表达特征；通过基因敲除和回补筛选出调控菌落相变的关键小RNA；通过gfp融合表达、凝胶阻滞等技术阐明其功能和调控方式，从而揭示小RNA在海洋病原弧菌环境适应性

中文关键词： 溶藻弧菌；小RNA；代谢；多糖；调控机制

英文摘要： Vibrio alginolyticus is one of the predominant pathogens of marine aquaculture in South China Sea. Previous study has found that the pathogen underwent colony phase variation between opaque and translucent. The two variants showed significant difference in the basic biochemical characteristics such as biofilm formation, motility and flagella biosynthesis, utilization of nitrogen and carbon resources, resistance to antibiotics and stress response as well. Genetic analysis has also revealed that complex signal transduction and genetic regulatory network including global regulators ValR, Rpos and ToxR are all involved in the phenomenon. However, the non-coding RNA (ncRNA) Hfq and associated small RNAs have been indicated to be the key factor in determination of the colony morphology. Deletion of hfq led to the change of the colony morphology, attenuation of the virulence, reduction of biofilm formation and stress responsibility, indicating critical roles of small RNAs in colony phase variation and associated cellular processes. To understand the mechanism, this project will first identify the small RNAs of V. alginolyticus and expression profiles by sequencing and comparing the transcriptoms of the wild type strains and the hfq mutants. The selected small RNAs will be confirmed and characterized by experimental app

英文关键词： vibrio alginolyticus；sRNA；metabolism；CPS；gene regulation