Background

Protein tags are protein or peptide sequences located either on the C- or N- terminal of the target protein, which facilitates one or several of the following characteristics: solubility, detection, purification, localization and expression. Maltose binding protein(MBP) is the 370 amino acid product of the E.coli mal E gene. MBP is a useful affinity tag that can increase the expression level and solubility of the resulting tagged protein. The MBP tag also promotes proper folding of the attached protein. 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 cross linked 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. An antibody to MBP can also be used to isolate or detect expression of the protein.