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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.

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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.


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.

[Indexed for MEDLINE]

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