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PMID: 19412171 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Automated unrestricted multigene recombineering for multiprotein complex production.

Nature methods ·Vol. 6 ·No. 6 ·2009-06-00 ·Pages 447-50

Bieniossek C, Nie Y, Frey D, Olieric N, Schaffitzel C, Collinson I, Romier C, Berger P, Richmond TJ, Steinmetz MO, Berger I

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.

MeSH Terms
Escherichia coli/physiology Multigene Family/genetics Protein Engineering/methods Recombinant Proteins/biosynthesis
Chemicals
Recombinant Proteins
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Bieniossek Christoph
European Molecular Biology Laboratory, Grenoble Outstation, Grenoble, France.
Nie Yan
Frey Daniel
Olieric Natacha
Schaffitzel Christiane
Collinson Ian
Romier Christophe
Berger Philipp
Richmond Timothy J
Steinmetz Michel O
Berger Imre
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14 references, click to expand
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Article Info
Journal
Nature methods
Abbr.
Nat Methods
ISSN
1548-7105
Published
2009-06-00
Epub
2009-00-03
Pages
447-50
Language
English
Region
United States
NLM ID
101215604
Subset
IM
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