Automated unrestricted multigene recombineering for multiprotein complex production.

Christoph Bieniossek (1, 2, 3) , Yan Nie (1, 2, 4) , Daniel Frey (5) , Natacha Olieric (5) , Christiane Schaffitzel (1, 2) , Ian Collinson (6) , Christophe Romier (7) , Philipp Berger (5) , Timothy J. Richmond (3) , Michel O. Steinmetz (5) , Imre Berger (1, 2)

1 European Molecular Biology Laboratory
2 Unit of Virus Host-Cell Interactions
3 Institut für Molekularbiologie und Biophysik [Zürich]
4 Department of Applied Physics
5 Biomolecular Research, Structural Biology
6 Department of Biochemistry
7 IGBMC - Institut de Génétique et de Biologie Moléculaire et Cellulaire

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.

Christoph Bieniossek, Yan Nie, Daniel Frey, Natacha Olieric, Christiane Schaffitzel, et al.. Automated unrestricted multigene recombineering for multiprotein complex production.. Nature Methods, 2009, 6 (6), pp.447-450. ⟨10.1038/nmeth.1326⟩. ⟨inserm-00420401⟩