title: Systems for the cell-free synthesis of proteins.
abstract: We describe a system for the cell-free expression of proteins based on extracts from Escherichia coli. Two reaction configurations, batch and continuous exchange, are discussed and analytical scale as well as preparative scale setups are documented. Guidelines for the systematic development and optimization of cell-free expression protocols are given in detail. We further provide specific protocols and parameters for the cell-free production of membrane proteins. High-throughput screening applications of CF expression systems are exemplified as new tools for genomics and proteomics studies.
authors.affiliation: Centre for Biomolecular Magnetic Resonance, Institute for Biophysical Chemistry, Goethe-University of Frankfurt/Main, Frankfurt/Main, Germany.
authors.forename: Lei
authors.initials: L
authors.lastname: Kai
authors.forename: Christian
authors.initials: C
authors.lastname: Roos
authors.forename: Stefan
authors.initials: S
authors.lastname: Haberstock
authors.forename: Davide
authors.initials: D
authors.lastname: Proverbio
authors.forename: Yi
authors.initials: Y
authors.lastname: Ma
authors.forename: Friederike
authors.initials: F
authors.lastname: Junge
authors.forename: Mikhail
authors.initials: M
authors.lastname: Karbyshev
authors.forename: Volker
authors.initials: V
authors.lastname: Dötsch
authors.forename: Frank
authors.initials: F
authors.lastname: Bernhard
mesh_terms: D017123:Bacteriophage T7 / Q000201:enzymology; D002458:Cell Fractionation; D004247:DNA / Q000235:genetics / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D003902:Detergents / Q000737:chemistry; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics*; D008274:Magnesium / Q000737:chemistry; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D003902:Detergents; D008565:Membrane Proteins; D011994:Recombinant Proteins; D004247:DNA; D012321:DNA-Directed RNA Polymerases; D008274:Magnesium