茶树油对SarA介导的金黄色葡萄球菌生物被膜形成的干预机制研究

项目名称： 茶树油对SarA介导的金黄色葡萄球菌生物被膜形成的干预机制研究

项目编号： No.31271951

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： 郭娜

作者单位： 吉林大学

项目金额： 86万元

中文摘要： 金黄色葡萄球菌（简称金葡菌）是一种常见食源性致病菌，具有形成生物被膜的能力。与浮游菌相比，被膜态细菌对防腐剂更具有耐性，更难从食品中清除。因此，研究致病菌生物被膜对食品加工和保藏具有重要意义。天然植物提取物- - 茶树油，作为一种潜在的防腐抗菌剂具有较好的干预金葡菌生物被膜形成的能力。目前由于该油抗生物被膜机制尚未清楚，限制了其作为食品防腐抗菌剂的开发应用。本项目拟首先借助生物芯片技术，分析茶树油压力下被膜态金葡菌的转录图谱，考察茶树油对转录调节因子SarA介导的生物被膜形成相关基因及信号通路的影响，揭示茶树油对SarA介导的金葡菌生物被膜形成的干预机制；再利用基因敲除技术、实时荧光定量PCR及免疫印迹技术验证芯片结果的正确性，运用激光扫描共聚焦显微成像系统和流式细胞术分析基因变化与表型的相关性。本研究将为茶树油的进一步开发应用提供理论依据，为其他天然食品防腐剂的抗菌机制研究提供新思路。

中文关键词： 茶树油；金黄色葡萄球菌生物被膜；分子机制；靶基因筛选；

英文摘要： Staphylococcus aureus is a common food-borne pathogenic bacterium. It has the ability to form biofilm. Bacteria living in a biofilm are more resistant to food preservatives and more difficult to remove from food compared to free-floating bacteria. Thus, research on pathogenic bacteria biofilm has important academic value in the fields of food processing and preservation. Natural plant extracts, tea tree oil, being a potential preservative, has the ability to disrupt the Staphylococcus aureus biofilm. At present, due to its resistance to biofilm mechanism has not yet clear. Because the mechanism of tea tree oil against the bacteria biofilm is not chear, its development and application as food preservatives was limited. In this program, we firstly plan to analyze the transcriptional profiles of Staphylococcus aureus grown in biofilm under tea tree oil stress using DNA microarray, in order to assay the the effect of tea tree oil on the genes and signal pathways of biofilm fomation mediated by SarA, a transcriptional regulator, and reveal the molecular mechanism of tea tree oil against the biofilm. To verify the microarray results by the technology such as knock-out, real time RT-PCR, western-blot. Moreover, flow cytometry and the laser confocal microscopy technique will be applied to analyse the relativity between

英文关键词： Tea Tree Oil；Staphylococcus aureus；molecular mechanism；target gene screening；