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

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

Author information

1
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.

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.

PMID:
25502194
DOI:
10.1007/978-1-4939-2230-7_4
[Indexed for MEDLINE]

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