Expression and purification of recombinant proteins by fusion to maltose-binding protein.
The pMAL vectors provide a method for purifying proteins from cloned genes by fusing them to maltose-binding protein (MBP, product of malE), which binds to amylose. The vectors use the tac promoter and the translation initiation signals of MBP to give high-level expression of the fusion, and an affinity purification for MBP to isolate the fusion protein. The pMAL polylinkers carry restriction sites to insert the gene of interest, and encode a site for a specific protease to separate MBP from the target protein after purification. Vectors with or without the malE signal sequence can be used, to express the protein cytoplasmically for the highest level of production or periplasmically to help in proper folding of disulfide-bonded proteins.
Riggs
P
P
New England Biolabs, Beverly, MA, USA. riggs@neb.com
D000688:Amylose / Q000737:chemistry; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D003001:Cloning, Molecular; D017886:Durapatite / Q000737:chemistry; D004765:Enteropeptidase / Q000235:genetics / Q000378:metabolism; D015951:Factor Xa / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D008320:Maltose / Q000302:isolation & purification; D058619:Maltose-Binding Proteins; D008969:Molecular Sequence Data; D010865:Pilot Projects; D011489:Protein Denaturation; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism
D002352:Carrier Proteins; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins; D008320:Maltose; D000688:Amylose; D017886:Durapatite; D015951:Factor Xa; D004765:Enteropeptidase