title: Flexible programming of cell-free protein synthesis using magnetic bead-immobilized plasmids.
abstract: The use of magnetic bead-immobilized DNA as movable template for cell-free protein synthesis has been investigated. Magnetic microbeads containing chemically conjugated plasmids were used to direct cell-free protein synthesis, so that protein generation could be readily programmed, reset and reprogrammed. Protein synthesis by using this approach could be ON/OFF-controlled through repeated addition and removal of the microbead-conjugated DNA and employed in sequential expression of different genes in a same reaction mixture. Since the incubation periods of individual template plasmids are freely controllable, relative expression levels of multiple proteins can be tuned to desired levels. We expect that the presented results will find wide application to the flexible design and execution of synthetic pathways in cell-free chassis.
authors.affiliation: Department of Fine Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon, Korea.
authors.forename: Ka-Young
authors.initials: KY
authors.lastname: Lee
authors.forename: Kyung-Ho
authors.initials: KH
authors.lastname: Lee
authors.forename: Ji-Woong
authors.initials: JW
authors.lastname: Park
authors.forename: Dong-Myung
authors.initials: DM
authors.lastname: Kim
mesh_terms: D002474:Cell-Free System; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008863:Microspheres; D010957:Plasmids / Q000378:metabolism*; D014176:Protein Biosynthesis
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins