DescriptionThe vector pMAL-p5X is designed to produce maltose-binding protein (MBP) fusions, where the protein of interest can be cleaved from MBP with the specific protease Factor Xa (NEB #P8010).
MBP fusions made with this vector include an N-terminal signal sequence, so the fusion protein is directed to the periplasm. The MBP has been engineered for tighter binding to amylose resin.
A gene or open reading frame is inserted into a restriction site of the vector polylinker, in the same translational reading frame as the malE gene (encoding maltose-binding protein). The fusion protein thus produced can be purified by amylose affinity chromatography. The sequence coding for the four amino acids Ile-Glu-Gly-Arg is present just upstream of the XmnI site. This allows the protein of interest to be cleaved from maltose-binding protein with the specific protease Factor Xa. Fragments inserted in the XmnI site (cleaves GAAGG↓ATTTC) will produce a fusion protein that, after Factor Xa cleavage, contains no vector-derived residues on the protein of interest.
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Properties and Usage
Affinity TagMaltose-Binding Protein (MBP)
Research Use Only
- The buyer/user has a non-exclusive license to use the vector for Research Purposes Only. Commercial use of this vector requires a license from New England Biolabs, Inc.
U.S. Patent No. 5,643,758
LicensesThe Buyer/User has a non-exclusive license to use this vector for RESEARCH PURPOSES ONLY. Commercial use requires a license from New England Biolabs, Inc. This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
- NEB 10-beta E. coli (High Efficiency) (NEB #C3019) is recommended for propagation and subcloning. NEB Express Competent E.coli (High Efficiency) (NEB #C2523) is recommended for expression using this vector.
- Guan, C., Li, P., Riggs, P.D. and Inouye, H. (1987). Gene. 67, 21-30.
- Maina, C.V., Riggs, P.D., Grandea, A.G.III, Slatko, B.E., Moran, L.S., Tagliamonte, J.A., McReynolds, L.A. and Guan, C. (1988). Gene. 74, 365-373.
- Nagai, K. and Thogersen, H.C. (1987). Methods Enzymology. 153, 461-481.
- Riggs, P.D. (1990). Expression and Purification of Maltose-Binding Protein Fusions. In F.M. Ausebel, R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith and K. Struhl (Ed.), Methods Enzymology. 16.6.1-16.6.12. New York: John Wiley & Sons, Inc.
- Yanisch-Perron, C., Vieira, J. and Messing, J. (1985). Gene. 33, 103-119.