Premium
A scalable, GFP‐based pipeline for membrane protein overexpression screening and purification
Author(s) -
Drew David,
Slotboom DirkJan,
Friso Giulia,
Reda Torsten,
Genevaux Pierre,
Rapp Mikaela,
MeindlBeinker Nadja M.,
Lambert Wietske,
Lerch Mirjam,
Daley Daniel O.,
Van Wijk KlaasJan,
Hirst Judy,
Kunji Edmund,
De Gier JanWillem
Publication year - 2005
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.051466205
Subject(s) - pipeline (software) , proteome , green fluorescent protein , escherichia coli , characterization (materials science) , protein purification , membrane protein , chemistry , computational biology , scalability , membrane , integral membrane protein , microbiology and biotechnology , biology , biochemistry , computer science , nanotechnology , gene , materials science , database , programming language
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.