Hultqvist Louise Dahl, Alhede Maria, Jakobsen Tim Holm, Givskov Michael, Bjarnsholt Thomas
Costerton Biofilm Centre, Department of Immunology and Microbiology, Faculty of Health Sciences, University of Copenhagen, Blegdamsvej 3B, 2200, Copenhagen, Denmark.
Singapore Centre on Environmental Life Sciences Engineering, Nanyang Technological University, Singapore, Singapore.
Methods Mol Biol. 2018;1673:203-212. doi: 10.1007/978-1-4939-7309-5_16.
In order to study N-acyl homoserine lactone (AHL)-based quorum sensing in vivo, we present a protocol using an Escherichia coli strain equipped with a luxR-based monitor system, which in the presence of exogenous AHL molecules expresses a green fluorescent protein (GFP). Lungs from mice challenged intratracheally with alginate beads containing both a Pseudomonas aeruginosa strain together with the E. coli monitor strain can be investigated at different time points postinfection. Epifluorescent or confocal scanning laser microscopy (CSLM) is used to detect the GFP-expressing E. coli monitor strain in the lung tissues, indicating production and excretion of AHLs in vivo by the infecting P. aeruginosa.
为了在体内研究基于N-酰基高丝氨酸内酯（AHL）的群体感应，我们提出了一种使用配备基于luxR监测系统的大肠杆菌菌株的方案，该系统在外源AHL分子存在时表达绿色荧光蛋白（GFP）。用含有铜绿假单胞菌菌株和大肠杆菌监测菌株的藻酸盐珠经气管内攻击小鼠后，可在感染后的不同时间点对肺进行研究。落射荧光显微镜或共聚焦扫描激光显微镜（CSLM）用于检测肺组织中表达GFP的大肠杆菌监测菌株，这表明感染的铜绿假单胞菌在体内产生并分泌了AHL。
