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

被引：105

作者：

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 ^{[1
]}

机构：

[1] Stockholm Univ, Dept Biochem & Biophys, SE-10691 Stockholm, Sweden

[2] MRC, Dunn Human Nutr Unit, Cambridge CB2 2XY, England

[3] Cornell Univ, Dept Plant Biol, Ithaca, NY 14853 USA

[4] Ctr Med, Dept Microbiol & Mol Med, CH-1211 Geneva, Switzerland

来源：

PROTEIN SCIENCE | 2005年 / 14卷 / 08期

基金：

英国医学研究理事会;

关键词：

Escherichia coli; membrane protein overexpression; membrane protein isolation; membrane protein characterization; GFP;

D O I：

10.1110/ps.051466205

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：2011 / 2017

页数：7

共 17 条

[1] Synthesis and bacterial expression of a gene encoding the heme domain of assimilatory nitrate reductase [J].