Chapter 2 - Detection and analysis of protein synthesis and RNA replication in giant liposomes

Abstract

In living cells, biochemical reaction systems are enclosed in small lipidic compartments. To experimentally simulate various biochemical reactions occurring in extant cells, giant liposomes are used to reconstruct an artificial model cell. We present methods for conducting a protein synthesis reaction, followed by the reaction catalyzed by the synthesized proteins inside liposomes, and for measurement of the in liposome reaction using a fluorescence-activated cell sorter (FACS). These techniques 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.