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Intra-Species Bacterial Quorum Sensing Studied at Single Cell Level in a Double Droplet Trapping System
Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK
Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QW, UK
Radboud University Nijmegen, Institute for Molecules and Materials, Heyendaalseweg 135, Nijmegen 6525 AJ, The Netherlands
These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 19 April 2013; in revised form: 9 May 2013 / Accepted: 10 May 2013 / Published: 21 May 2013
Abstract: 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.
Keywords: droplet-based microfluidics; bacterial quorum sensing; single cell
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Bai, Y.; Patil, S.N.; Bowden, S.D.; Poulter, S.; Pan, J.; Salmond, G.P.C.; Welch, M.; Huck, W.T.S.; Abell, C. Intra-Species Bacterial Quorum Sensing Studied at Single Cell Level in a Double Droplet Trapping System. Int. J. Mol. Sci. 2013, 14, 10570-10581.
Bai Y, Patil SN, Bowden SD, Poulter S, Pan J, Salmond GPC, Welch M, Huck WTS, Abell C. Intra-Species Bacterial Quorum Sensing Studied at Single Cell Level in a Double Droplet Trapping System. International Journal of Molecular Sciences. 2013; 14(5):10570-10581.
Bai, Yunpeng; Patil, Santoshkumar N.; Bowden, Steven D.; Poulter, Simon; Pan, Jie; Salmond, George P.C.; Welch, Martin; Huck, Wilhelm T.S.; Abell, Chris. 2013. "Intra-Species Bacterial Quorum Sensing Studied at Single Cell Level in a Double Droplet Trapping System." Int. J. Mol. Sci. 14, no. 5: 10570-10581.