Characterization and production of protein complexes by co-expression in Escherichia coli

Methods Mol Biol. 2015;1261:63-89. doi: 10.1007/978-1-4939-2230-7_4.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular / methods*
  • DNA-Binding Proteins / biosynthesis
  • DNA-Binding Proteins / genetics
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism
  • Multiprotein Complexes / biosynthesis*
  • Multiprotein Complexes / genetics
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics

Substances

  • DNA-Binding Proteins
  • Multiprotein Complexes
  • Recombinant Proteins