Bai Yunpeng, Patil Santoshkumar N, Bowden Steven D, Poulter Simon, Pan Jie, Salmond George P C, Welch Martin, Huck Wilhelm T S, Abell Chris
Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK.
Int J Mol Sci. 2013 May 21;14(5):10570-81. doi: 10.3390/ijms140510570.
In this paper, we investigated the intra-species bacterial quorum sensing at the single cell level using a double droplet trapping system. Escherichia coli transformed to express the quorum sensing receptor protein, LasR, were encapsulated in microdroplets that were positioned adjacent to microdroplets containing the autoinducer, N-(3-oxododecanoyl)-L-homoserine lactone (OdDHL). Functional activation of the LasR protein by diffusion of the OdDHL across the droplet interface was measured by monitoring the expression of green fluorescent protein (GFP) from a LasR-dependent promoter. A threshold concentration of OdDHL was found to induce production of quorum-sensing associated GFP by E. coli. Additionally, we demonstrated that LasR-dependent activation of GFP expression was also initiated when the adjacent droplets contained single E. coli transformed with the OdDHL synthase gene, LasI, representing a simple quorum sensing circuit between two droplets.
在本文中，我们使用双液滴捕获系统在单细胞水平上研究了种内细菌群体感应。将转化后表达群体感应受体蛋白LasR的大肠杆菌封装在微滴中，这些微滴与含有自诱导剂N-(3-氧代十二烷酰基)-L-高丝氨酸内酯(OdDHL)的微滴相邻放置。通过监测来自LasR依赖性启动子的绿色荧光蛋白(GFP)的表达，来测量OdDHL通过液滴界面扩散对LasR蛋白的功能激活。发现OdDHL的阈值浓度可诱导大肠杆菌产生群体感应相关的GFP。此外，我们还证明，当相邻液滴包含用OdDHL合酶基因LasI转化的单个大肠杆菌时，也会启动LasR依赖性的GFP表达激活，这代表了两个液滴之间的一个简单群体感应回路。
