Automated unrestricted multigene recombineering for multiprotein complex production.

Bieniossek, Christoph; Nie, Yan; Frey, Daniel; Olieric, Natacha; Schaffitzel, Christiane; Collinson, Ian; Romier, Christophe; Berger, Philipp et al. · Nat Methods · 2009

basic_science · Level V

Where this comes from

Abstract

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.

Medical subject headings