title: Protein synthesis in liposomes with a minimal set of enzymes.
abstract: In a significant step towards the construction of the semi-synthetic minimal cell, a protein expression system with a minimal set of pure and specific enzymes is required. A novel cell-free transcription and translation system named PURESYSTEM (PS), consisting of a specified set of 36 enzymes and ribosomes, has been entrapped in POPC liposomes for protein synthesis. The PS has been used to transcribe and translate an Enhanced Green Fluorescent Protein (EGFP) gene from plasmid DNA. The synthesis is confirmed by the EGFP fluorescence emitting liposomes on fluorometric analysis and on confocal microscopy analysis. Furthermore the PS encapsulated into POPC liposomes can drive the expression of the plsB and plsC genes encoding for the sn-glycerol-3-phosphate acyltransferase (GPAT) and 1-acyl-sn-glycerol-3-phosphate acyltransferase (LPAAT) involved in the first step of the "salvage pathway" for synthesis of POPC. The expression of GPAT and LPAAT in liposomes would in principle allow the production of the cell boundary from within.
authors.affiliation: Centro Studi e Ricerche E. Fermi, Compendio Viminale, 00184 Rome, Italy. murtas@uniroma3.it
authors.forename: Giovanni
authors.initials: G
authors.lastname: Murtas
authors.forename: Yutetsu
authors.initials: Y
authors.lastname: Kuruma
authors.forename: Paolo
authors.initials: P
authors.lastname: Bianchini
authors.forename: Alberto
authors.initials: A
authors.lastname: Diaspro
authors.forename: Pier Luigi
authors.initials: PL
authors.lastname: Luisi
mesh_terms: D032701:Biomimetics / Q000379:methods*; D002474:Cell-Free System; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry*; D009097:Multienzyme Complexes / Q000737:chemistry*; D011506:Proteins / Q000138:chemical synthesis*
chemical_list: D029968:Escherichia coli Proteins; D008081:Liposomes; D009097:Multienzyme Complexes; D011506:Proteins