Automated unrestricted multigene recombineering for multiprotein complex production

Christoph Bieniossek, Yan Nie, Daniel Frey, Natacha Olieric, Christiane Schaffitzel, Ian Collinson, Christophe Romier, Philipp Berger, Timothy J Richmond, Michel O Steinmetz, Imre Berger

Research output: Contribution to journalArticle (Academic Journal)peer-review

74 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)447 - 450
Number of pages4
JournalNature Methods
Volume6
Issue number6
DOIs
Publication statusPublished - Jun 2009

Bibliographical note

Other: First published online 3rd May 2009

Keywords

  • Escherichia coli
  • Multigene Family
  • Protein Engineering
  • Recombinant Proteins

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