[Expression, export and one-step purification of proteins by fusion to the MalE protein of E. coli].
Enzymes can be fused at the C-terminal end of the maltose binding protein (MalE), at the genetic level. Expression of the hybrid proteins, under control of promoter malEp and of the constitutive activator, MalTc1, can be repressed by glucose. The hybrid proteins are localised either in the bacterial cytoplasm or periplasmic space, depending on whether MalE harbors a signal peptide mutation or not; as MalE, they can be purified in one step by chromatography on cross-linked amylose. The Staphylococcus aureus Nuclease and the Klenow portion of E. coli DNA-polymerase I keep their specific activities when fused to MalE.
D018528:ATP-Binding Cassette Transporters; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002846:Chromatography, Affinity / Q000379:methods; D004798:Enzymes / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D033902:Periplasmic Binding Proteins; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*
D018528:ATP-Binding Cassette Transporters; D002352:Carrier Proteins; D004798:Enzymes; D029968:Escherichia coli Proteins; C461619:MalE protein, E coli; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D033902:Periplasmic Binding Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C461599:maltose transport system, E coli
Expression, exportation et purification en une étape de protéines par fusion à la protéine MalE d'E. coli.