Faculty Opinions recommendation of A vesicle bioreactor as a step toward an artificial cell assembly.
Year: 2005
Type: peer-review
Abstract: An Escherichia coli cell-free expression system is encapsulated in a phospholipid vesicle to build a cell-like bioreactor.Large unilamellar vesicles containing extracts are produced in an oil-extract emulsion.To form a bilayer the vesicles are transferred into a feeding solution that contains ribonucleotides and amino acids.Transcription-translation of plasmid genes is isolated in the vesicles.Whereas in bulk solution expression of enhanced GFP stops after 2 h, inside the vesicle permeability of the membrane to the feeding solution prolongs the expression for up to 5 h.To solve the energy and material limitations and increase the capacity of the reactor, the ␣-hemolysin pore protein from Staphylococcus aureus is expressed inside the vesicle to create a selective permeability for nutrients.The reactor can then sustain expression for up to 4 days with a protein production of 30 M after 4 days.Oxygen diffusion and osmotic pressure are critical parameters to maintain expression and avoid vesicle burst. ␣-hemolysin ͉ cell-free protein expression ͉ membrane-anchoring polypeptide
Author: John Glass
Institution: J. Craig Venter Institute
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Funder: Charles H. Revson Foundation
OpenAlex ID: W4210924561