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

doi:10.1038/nmeth.1326

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

School of Biochemistry

Research output: Contribution to journal › Article (Academic Journal) › peer-review

81 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 language	English
Pages (from-to)	447 - 450
Number of pages	4
Journal	Nature Methods
Volume	6
Issue number	6
DOIs	https://doi.org/10.1038/nmeth.1326
Publication status	Published - Jun 2009

Bibliographical note

Other: First published online 3rd May 2009

Keywords

Escherichia coli
Multigene Family
Protein Engineering
Recombinant Proteins

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title = "Automated unrestricted multigene recombineering for multiprotein complex production",

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.",

keywords = "Escherichia coli, Multigene Family, Protein Engineering, Recombinant Proteins",

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

note = "Other: First published online 3rd May 2009",

year = "2009",

month = jun,

doi = "10.1038/nmeth.1326",

language = "English",

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T1 - Automated unrestricted multigene recombineering for multiprotein complex production

AU - Bieniossek, Christoph

AU - Nie, Yan

AU - Frey, Daniel

AU - Olieric, Natacha

AU - Schaffitzel, Christiane

AU - Collinson, Ian

AU - Romier, Christophe

AU - Berger, Philipp

AU - Richmond, Timothy J

AU - Steinmetz, Michel O

AU - Berger, Imre

N1 - Other: First published online 3rd May 2009

PY - 2009/6

Y1 - 2009/6

N2 - 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.

AB - 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.

KW - Escherichia coli

KW - Multigene Family

KW - Protein Engineering

KW - Recombinant Proteins

U2 - 10.1038/nmeth.1326

DO - 10.1038/nmeth.1326

M3 - Article (Academic Journal)

C2 - 19412171

VL - 6

SP - 447

EP - 450

JO - Nature Methods

JF - Nature Methods

SN - 1548-7091

IS - 6

ER -

Automated unrestricted multigene recombineering for multiprotein complex production

Abstract

Bibliographical note

Keywords

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