Journal
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
Volume 118, Issue 2, Pages 199-204Publisher
SOC BIOSCIENCE BIOENGINEERING JAPAN
DOI: 10.1016/j.jbiosc.2014.01.014
Keywords
Cell-free protein synthesis; Microcompartmentalization; Semipermeable microcapsule; Microfluidics; Rupture-induced encapsulation
Funding
- Nanotechnology Network Japan project of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan
- MEXT
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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. (C) 2014, The Society for Biotechnology, Japan. All rights reserved.
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