title: Cell-Free Synthesis of Macromolecular Complexes.
abstract: Cell-free protein synthesis based on E. coli cell extracts has been described for the first time more than 50 years ago. To date, cell-free synthesis is widely used for the preparation of toxic proteins, for studies of the translation process and its regulation as well as for the incorporation of artificial or labeled amino acids into a polypeptide chain. Many efforts have been directed towards establishing cell-free expression as a standard method for gene expression, with limited success. In this chapter we will describe the state-of-the-art of cell-free expression, extract preparation methods and recent examples for successful applications of cell-free synthesis of macromolecular complexes.
authors.affiliation: European Molecular Biology Laboratory, Grenoble Outstation, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Botte
authors.affiliation: European Molecular Biology Laboratory, Grenoble Outstation, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France. aurelien.deniaud@cea.fr.|CEA Grenoble, iRTSV/LCBM/BioMet, 17 rue des Martyrs, 38054, Grenoble, France. aurelien.deniaud@cea.fr.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Deniaud
authors.affiliation: European Molecular Biology Laboratory, Grenoble Outstation, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France. schaffitzel@embl.fr.|School of Biochemistry, University of Bristol, Bristol, BS8 1TD, UK. schaffitzel@embl.fr.
authors.forename: Christiane
authors.initials: C
authors.lastname: Schaffitzel
mesh_terms: D000818:Animals; D002474:Cell-Free System; D005786:Gene Expression Regulation; D006801:Humans; D008958:Models, Molecular; D046912:Multiprotein Complexes; D015202:Protein Engineering / Q000379:methods*; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D012270:Ribosomes / Q000378:metabolism; D013329:Structure-Activity Relationship; D014158:Transcription, Genetic
chemical_list: D046912:Multiprotein Complexes; D021122:Protein Subunits; D011994:Recombinant Proteins
keywords: Cell-free synthesis; Membrane proteins; Ribosomes; Synthetic biology