16 May 2009
Nat Methods. 2009 May 3. [Epub ahead of print]
Automated unrestricted multigene recombineering for multiprotein complex production.
Bieniossek C, Nie Y, Frey D, Olieric N, Schaffitzel C, Collinson I, Romier C, Berger P, Richmond TJ, Steinmetz MO,Berger I.
[1] European Molecular Biology Laboratory, Grenoble Outstation, B.P. 181, Grenoble, France. [2] Unit of Virus Host-Cell Interactions, Unités Mixtes de Recherche 5233, Grenoble, France. [3] Eidgenössische Technische Hochschule Zürich, Institut für Molekularbiologie und Biophysik, Hönggerberg, Zürich, Switzerland. [4] 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.
