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PMID: 15987891 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A scalable, GFP-based pipeline for membrane protein overexpression screening and purification.

Protein science : a publication of the Protein Society ·Vol. 14 ·No. 8 ·2005-08-00 ·Pages 2011-7

Drew D, Slotboom DJ, Friso G, Reda T, Genevaux P, Rapp M, Meindl-Beinker NM, Lambert W, Lerch M, Daley DO, Van Wijk KJ, Hirst J, Kunji E, De Gier JW

Abstract

We describe a generic, GFP-based pipeline for membrane protein overexpression and purification in Escherichia coli. We exemplify the use of the pipeline by the identification and characterization of E. coli YedZ, a new, membrane-integral flavocytochrome. The approach is scalable and suitable for high-throughput applications. The GFP-based pipeline will facilitate the characterization of the E. coli membrane proteome and serves as an important reference for the characterization of other membrane proteomes.

MeSH Terms
Escherichia coli/genetics Escherichia coli Proteins/biosynthesis,genetics,isolation & purification Green Fluorescent Proteins/analysis,genetics Lactococcus lactis/genetics Luminescent Agents/analysis Membrane Proteins/biosynthesis,genetics,isolation & purification Recombinant Fusion Proteins/biosynthesis,isolation & purification
Chemicals
Escherichia coli Proteins Luminescent Agents Membrane Proteins Recombinant Fusion Proteins Green Fluorescent Proteins
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
Drew David
Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
Slotboom Dirk-Jan
Friso Giulia
Reda Torsten
Genevaux Pierre
Rapp Mikaela
Meindl-Beinker Nadja M
Lambert Wietske
Lerch Mirjam
Daley Daniel O
Van Wijk Klaas-Jan
Hirst Judy
Kunji Edmund
De Gier Jan-Willem
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Article Info
Journal
Protein science : a publication of the Protein Society
Abbr.
Protein Sci
ISSN
0961-8368
Published
2005-08-00
Epub
2005-00-29
Pages
2011-7
Language
English
Region
United States
NLM ID
9211750
PMCID
PMC2279312
Subset
IM
Grants
Medical Research Council · MC_U105663139 · United Kingdom
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