Antibody / Maltose Binding Protein / malE

Plasmid vectors for the expression of coding regions of eukaryotic genes in bacterial, insect and mammalian hosts are in common usage; such expression vectors frequently encode hybrid fusion proteins consisting in part of prokaryotic and in part, eukaryotic specified proteins. One such system utilizes maltose binding protein (MBP), the 370 amino acid product of the E. coli mal E gene. Plasmid vectors have been constructed utilizing the MBP domain that allow the synthesis of high levels of MBP-fusion proteins that can be Purified in a one step procedure by affinity chromatography crosslinked amylose resin. Once bound to amylose, the MBP protein can then be separated from the target protein by cleavage by coagulation factor Xa at a specific four residue site. Alternatively, the intact fusion protein can be specifically eluted from the resin by the addition of excess free maltose. Subsequent to elution, MBP fusion protein can be visualized either by Western Blot Analysis or immunoprecipitation using antibodies specific for the MBP-tag. Expression systems utilizing the MBP fusion tag include pCG-806fx and pMal vectors.