Brief Communication abstract
Nature Methods 6, 447 - 450 (2009)
Published online: 3 May 2009 | doi:10.1038/nmeth.1326
Automated unrestricted multigene recombineering for multiprotein complex production
Christoph Bieniossek1,2,3,8, Yan Nie1,2,4,8, Daniel Frey5, Natacha Olieric5, Christiane Schaffitzel1,2, Ian Collinson6, Christophe Romier7, Philipp Berger5, Timothy J Richmond3, Michel O Steinmetz5 & Imre Berger1,2
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.
- 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.
- Department of Applied Physics, Royal Institute of Technology, Albanova University Center, Stockholm, Sweden.
- Biomolecular Research, Structural Biology, Paul Scherrer Institut, Villigen, Switzerland.
- Department of Biochemistry, School of Medical Sciences, Bristol, UK.
- Department of Biology and Structural Genomics, Institute Génétique et Biologie Moléculaire et Cellulaire, Illkirch, France.
- These authors contributed equally to this work.
Correspondence to: Michel O Steinmetz5 e-mail: michel.steinmetz@psi.ch
Correspondence to: Imre Berger1,2 e-mail: iberger@embl.fr
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