title: Expression of membrane-associated proteins within single emulsion cell facsimiles.
abstract: MreB is a structural membrane-associated protein which is one of the key components of the bacterial cytoskeleton. Although it plays an important role in shape maintenance of rod-like bacteria, the understanding of its mechanism of action is still not fully understood. This study shows how segmented flow and microdroplet technology can be used as a new tool for biological in vitro investigation of this protein. In this paper, we demonstrate cell-free expression in a single emulsion system to express red fluorescence protein (RFP) and MreB linked RFP (MreB-RFP). We follow the aggregation and localisation of the fusion protein MreB-RFP in this artificial cell-like environment. The expression of MreB-RFP in single emulsion droplets leads to the formation of micrometer-scale protein patches distributed at the water/oil interface.
authors.affiliation: The Division of Biomedical Engineering, School of Engineering, The University of Glasgow, G12 8LT, Glasgow, UK.
authors.forename: Mayuree
authors.initials: M
authors.lastname: Chanasakulniyom
authors.forename: Chiara
authors.initials: C
authors.lastname: Martino
authors.forename: David
authors.initials: D
authors.lastname: Paterson
authors.forename: Louise
authors.initials: L
authors.lastname: Horsfall
authors.forename: Susan
authors.initials: S
authors.lastname: Rosser
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Cooper
mesh_terms: D002474:Cell-Free System; D004655:Emulsions; D008565:Membrane Proteins / Q000378:metabolism*; D044085:Microfluidics; D018613:Microscopy, Confocal; D010957:Plasmids
chemical_list: D004655:Emulsions; D008565:Membrane Proteins