title: Characterization and production of protein complexes by co-expression in Escherichia coli.
abstract: The functional units within cells are often macromolecular complexes rather than single species. Production of these complexes as assembled homogenous samples is a prerequisite for their biophysical and structural characterization and hence an understanding of their function in molecular terms. Co-expression in Escherichia coli has been used routinely to decipher the subunit composition, assembly, and production of whole protein complexes. Such complexes can then be used to reconstitute protein/nucleic acid complexes in vitro. In this chapter we present protocols for the widely utilized ACEMBL and pET-MCN/pET-MCP vector series which enable the rapid and automated co-expression of protein complexes in Escherichia coli.
authors.affiliation: European Molecular Biology Laboratory (EMBL), Grenoble Outstation and Unit of Virus Host-Cell Interactions (UVHCI), Université Grenoble Alpes-EMBL-CNRS, UMR 5233, 6 rue Jules Horowitz, 38042, Grenoble Cedex 9, France.
authors.forename: Matthias
authors.initials: M
authors.lastname: Haffke
authors.forename: Martin
authors.initials: M
authors.lastname: Marek
authors.forename: Martin
authors.initials: M
authors.lastname: Pelosse
authors.forename: Marie-Laure
authors.initials: ML
authors.lastname: Diebold
authors.forename: Uwe
authors.initials: U
authors.lastname: Schlattner
authors.forename: Imre
authors.initials: I
authors.lastname: Berger
authors.forename: Christophe
authors.initials: C
authors.lastname: Romier
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D046912:Multiprotein Complexes / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D004268:DNA-Binding Proteins; D046912:Multiprotein Complexes; D011994:Recombinant Proteins