title: Cell-free protein synthesis for functional and structural studies.
abstract: Recent advances in cell-free protein expression systems have made them reliable and practical for functional and structural studies of a wide variety of proteins. In particular, wheat germ cell-free translation can consistently produce target proteins in microgram quantities from relatively inexpensive, small-scale reactions. Here we describe our small-scale protein expression method for rapidly producing proteins for functional assay and techniques for determining if the target is suitable for scale-up to amounts potentially needed for structure determination. The cell-free system is versatile and can be easily customized with the inclusion of additives. We describe simple modifications used for producing membrane proteins.
authors.affiliation: Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Makino
authors.forename: Emily T
authors.initials: ET
authors.lastname: Beebe
authors.forename: John L
authors.initials: JL
authors.lastname: Markley
authors.forename: Brian G
authors.initials: BG
authors.lastname: Fox
mesh_terms: D002474:Cell-Free System; D003956:Dialysis; D019286:Endopeptidase K / Q000378:metabolism; D008081:Liposomes; D008565:Membrane Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D010940:Plant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility
chemical_list: D008081:Liposomes; D008565:Membrane Proteins; D010940:Plant Proteins; D011506:Proteins; D019286:Endopeptidase K