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Description:
The vector pMAL-p2G is designed to produce maltose-binding protein (MBP) fusions, where the protein of interest can be cleaved from MBP with the specific protease Genenase™ I (NEB #P8075). The malE gene on this vector includes the sequence coding for the amino-terminal signal peptide of MBP, so the fusion protein is directed to the periplasm of E. coli.
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 MBP). Insertion of the DNA fragment interrupts the malE-lacZ α fusion preexisting on the vector, affording a screen for inserts on the proper indicator plates. The fusion protein produced from the vector can be purified by amylose affinity chromatography. The sequence coding for the six amino acids Pro-Gly-Ala-Ala-His-Tyr is present just upstream of the SnaBI site. This allows the protein of interest to be cleaved from MBP with the specific protease Genenase™ I.
pMAL-p2G cut with SnaBI produces a blunt end at the tyrosine codon. This allows blunt-end cloning of an insert where the first three nucleotides code for the first amino acid of the protein of interest, and Genenase™ I cleavage of the fusion produces a protein with no vector-derived amino acids.
Source:
ER2272 (pMAL-p2G)
Storage Conditions
Storage Temperature:
-20°C
References
- Guan, C., Li, P., Riggs, P.D. and Inouye, H. (1988) Vectors that facilitate the expression and purification of foreign peptides in Escherichia coli by fusion to maltose-binding protein. 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) R. Wu and L. Grossman (Eds.), Methods Enzymol., 153, pp. 461-481. San Diego: Academic Press.
- Riggs, P.D. (1990) In Expression and Purification of Maltose-Binding Protein Fusions. F.M. Ausubel, R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith and K. Struhl (Eds.), Current Protocols in Molecular Biology, pp. 16.6.1-16.6.12.
- Carter, P. et al. (1989) Proteins: Structure, Function, and Genetics, 6, 240-248.
Companion Products
pMAL Protein Fusion and Purification System
pMAL-c2E Vector
Legal
Research Use Assurance:
The Buyer/User has a non-exclusive license to use the pMAL-p2G vector for RESEARCH PURPOSES ONLY. For commercial use of this vector, both Non-profit and For-profit buyers and users may obtain a license from New England Biolabs, Inc. U.S. Patent No. 5,643,758.
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