European Molecular Biology Laboratory, Grenoble Outstation, B.P. 181, Grenoble, France.  Unit of Virus Host-Cell Interactions, Unités Mixtes de Recherche 5233, Grenoble, France.  Eidgenössische Technische Hochschule Zürich, Institut für Molekularbiologie und Biophysik, Hönggerberg, Zürich, Switzerland.  These authors contributed equally to this work.
Structural and functional studies of many multiprotein complexes depend on recombinant-protein overexpression. Rapid revision of expression experiments and diversification of the complexes are often crucial for success of these projects; therefore, automation is increasingly indispensable. We introduce Acembl, a versatile and automatable system for protein-complex expression in Escherichia coli that uses recombineering to facilitate multigene assembly and diversification. We demonstrated protein-complex expression using Acembl, including production of the complete prokaryotic holotranslocon.