title: Microcompartmentalized cell-free protein synthesis in semipermeable microcapsules composed of polyethylenimine-coated alginate.
abstract: We describe microcompartmentalized cell-free protein synthesis in semipermeable microcapsules prepared from water-in-oil-in-water droplets by a rupture-induced encapsulation method. An aqueous solution of template DNA coding for green fluorescent protein and enzymes for the cell-free protein synthesis was aliquoted into water-in-oil droplets using a microfluidic device, and the droplets were transformed into semipermeable microcapsules. Substrates for protein synthesis diffused into the microcapsules through their semipermeable polyion complex membranes composed of polyethylenimine-coated alginate. Cell-free protein synthesis was confirmed by detection of the fluorescence of the synthesized green fluorescence protein in the microcapsules. We also used this microcompartmentalized system to synthesize protein from a single molecule of template DNA encapsulated by limiting dilution.
authors.affiliation: Faculty of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan; Department of Chemical Science and Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo 657-8501, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Saeki
authors.affiliation: National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan. Electronic address: shinji.sugiura@aist.go.jp.
authors.forename: Shinji
authors.initials: S
authors.lastname: Sugiura
authors.affiliation: National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Kanamori
authors.affiliation: Faculty of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan.
authors.forename: Seigo
authors.initials: S
authors.lastname: Sato
authors.affiliation: Faculty of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan. Electronic address: ichikawa.sosaku.fn@u.tsukuba.ac.jp.
authors.forename: Sosaku
authors.initials: S
authors.lastname: Ichikawa
mesh_terms: D000464:Alginates / Q000737:chemistry*; D002214:Capsules / Q000737:chemistry*; D004247:DNA / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D046210:Microfluidic Analytical Techniques; D010539:Permeability; D011094:Polyethyleneimine / Q000737:chemistry*; D014176:Protein Biosynthesis
chemical_list: D000464:Alginates; D002214:Capsules; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine; D004247:DNA
keywords: Cell-free protein synthesis; Microcompartmentalization; Microfluidics; Rupture-induced encapsulation; Semipermeable microcapsule