In living cells, biochemical reaction systems are enclosed in the small lipidic compartments. To experimentally simulate various biochemical reactions occurring in extant cells, cell-sized lipid vesicles (liposomes) are used to reconstruct an artificial model cell. We present methods for encapsulation of the protein synthesis system inside liposomes and for measurement of the in liposome synthesis reaction using a fluorescence-activated cell sorter. These techniques would enable us to perform detailed analysis of the biochemical reactions occurring in the microcompartments and have the potential to reveal the role of compartmentalization in cellular systems.
Affiliation(s): (1) Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, Osaka, Japan
Series: Methods in Molecular Biology | Volume: 607 | Pub. Date: Aug-01-2009 | Page Range: 243-256 | DOI: 10.1007/978-1-60327-331-2_20
Subject: Protein Science
Key Words: Liposome - Flow cytometry - Fluorescence-activated cell sorter - Cell-free protein synthesis - Green fluorescent protein - β-Galactosidase - β-Glucuronidase