沙门氏菌生物菌膜中特征性非编码小RNA的鉴别及调控机制

项目名称： 沙门氏菌生物菌膜中特征性非编码小RNA的鉴别及调控机制

项目编号： No.31501574

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 农业科学

项目作者： 王虎虎

作者单位： 南京农业大学

项目金额： 20万元

中文摘要： 生物菌膜是食品加工中沙门氏菌污染的重要传播根源，其与食物中毒和食品召回有密切关联，探明食品加工环境中生物菌膜的形成机制非常必要；非编码小RNA（Non-coding small RNA，sRNAs）是一类新型的细菌调控因子，前期研究发现sRNAs参与在沙门氏菌生物菌膜的成熟调控中，但何种标志性sRNAs发挥着关键调控作用，以及具体的调控途径均不清楚；本研究拟以肠炎沙门氏菌为对象，以食品级不锈钢为接触面，以肉汁液（meat thawing-loss broth，MTLB）和标准培养液（TSB）为生长基质对比研究，利用RNA-seq技术结合生物信息学挖掘生物菌膜中的特征性sRNAs（3-6个），在探明其序列位点和保守性的基础上，通过构建特征性sRNAs的缺失突变株，结合粘附表型差异，揭示sRNAs对生物菌膜的调控机制；该成果可完善生物菌膜的科学理论，并为研发生物菌膜的防控技术奠定理论基础。

中文关键词： 生物膜；沙门氏菌；高通量筛选；非编码小RNA；肉汁液

英文摘要： Biofilm is the main contamination source of Salmonella spread in food-processing industry, Salmonella biofilm has been directly associated with many food-borne outbreaks and food recalls, it was very essential to reveal the mechanism of Salmonella biofilm formation under food processing environments. Non-coding small RNA (sRNAs) was found as a new regulatory element in bacteria. Previous studies have found that several sRNAs have involved in regulation of maturation of Salmonella biofilm, However, which characteristic sRNAs played key roles in biofilm formation, and the concrete regulation pathways were still unknown. In the present study, Salmonella Enteritidis was tested as the target bacteria, and the food-grade stainless steel surface was used for biofilm formation. As compared with a standard growth medium (TSB), a meat thawing-loss broth (MTLB) was used for Salmonella growth. The novel sRNAs (3-6) regulating biofilm formation of Salmonella in MTLB were genome-wide identified with RNA-seq and bioinformatics technologies. On the basis of confirming the sequences sites and conservation, the molecular regulation mechanisms of target sRNAs in biofilm formation were clarified by constructing the deleted mutant strains of novel sRNAs and analyzing the differences of adhesion phenotypes between mutants and wild-type Salmonella. Our study will perfect the scientific theory of biofilm formation, and also establish the theoretical foundation for developing effective prevention and control measures of Salmonella biofilm.

英文关键词： Biofilm;Salmonella;High-throughput screening;Non-coding small RNA;Meat-based broth