An Escherichia coli vector to express and purify foreign proteins by fusion to and separation from maltose-binding protein.
A plasmid vector has been constructed that directs the synthesis of high levels (approximately 2% of total cellular protein) of fusions between a target protein and maltose-binding protein (MBP) in Escherichia coli. The MBP domain is used to purify the fusion protein in a one step procedure by affinity chromatography to crosslinked amylose resin. The fusion protein contains the recognition sequence (Ile-Glu-Gly-Arg) for blood coagulation factor Xa protease between the two domains. Cleavage by factor Xa separates the two domains and the target protein domain can then be purified away from the MBP domain by repeating the affinity chromatography step. A prokaryotic (beta-galactosidase) and a eukaryotic (paramyosin) protein have been successfully purified by this method.
Maina
C V
CV
New England Biolabs, Beverly, MA 01915.
Riggs
P D
PD
Grandea
A G
AG
Slatko
B E
BE
Moran
L S
LS
Tagliamonte
J A
JA
McReynolds
L A
LA
Guan
C D
CD
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins / Q000235:genetics*; D002352:Carrier Proteins / Q000235:genetics*; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D005822:Genetic Vectors; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000235:genetics*
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C461599:maltose transport system, E coli