PMID:26213264

Citation	Jung, Y, Jung, H and Lim, D (2015) Bacteriophage membrane protein P9 as a fusion partner for the efficient expression of membrane proteins in Escherichia coli. Protein Expr. Purif. 116:12-8
Abstract	Despite their important roles and economic values, studies of membrane proteins have been hampered by the difficulties associated with obtaining sufficient amounts of protein. Here, we report a novel membrane protein expression system that uses the major envelope protein (P9) of phage φ6 as an N-terminal fusion partner. Phage membrane protein P9 facilitated the synthesis of target proteins and their integration into the Escherichia coli cell membrane. This system was used to produce various multi-pass transmembrane proteins, including G-protein-coupled receptors, transporters, and ion channels of human origin. Green fluorescent protein fusion was used to confirm the correct folding of the expressed proteins. Of the 14 membrane proteins tested, eight were highly expressed, three were moderately expressed, and three were barely expressed in E. coli. Seven of the eight highly expressed proteins could be purified after extraction with the mild detergent lauryldimethylamine-oxide. Although a few proteins have previously been developed as fusion partners to augment membrane protein production, we believe that the major envelope protein P9 described here is better suited to the efficient expression of eukaryotic transmembrane proteins in E. coli.
Links	PubMed Online version:10.1016/j.pep.2015.07.010
Keywords	Amino Acid Sequence; Bacteriophage phi 6/chemistry; Bacteriophage phi 6/genetics; Base Sequence; Cloning, Molecular; Detergents/chemistry; Escherichia coli/genetics; Humans; Ion Channels/chemistry; Ion Channels/genetics; Ion Channels/isolation & purification; Molecular Sequence Data; Plasmids/chemistry; Plasmids/genetics; Receptors, G-Protein-Coupled/chemistry; Receptors, G-Protein-Coupled/genetics; Receptors, G-Protein-Coupled/isolation & purification; Recombinant Fusion Proteins/chemistry; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/isolation & purification; Up-Regulation; Viral Proteins/chemistry; Viral Proteins/genetics; Viral Proteins/isolation & purification
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
BPPH6:P9		GO:0044385: integral to membrane of host cell	ECO:0000314:		C		organism: phage phi6 protein P9//P9 Figure 1D uses immunoblot to show the localization of the P9 in the cells using sonication.	complete CACAO 12872

References

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