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

Fiche publication


Date publication

janvier 2015

Auteurs

Membres identifiés du Cancéropôle Est :
Dr ROMIER Christophe


Tous les auteurs :
Haffke M, Marek M, Pelosse M, Diebold ML, Schlattner U, Berger I, Romier C

Résumé

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.

Référence

Methods Mol Biol. 2015;1261:63-89