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Description:
The vector pMAL-p2E is designed to produce maltose-binding protein (MBP) fusions, where the protein of interest can be cleaved from MBP with the specific protease enterokinase
(NEB #P8070).
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.
Source:
ER2272 (pMAL-p2E)
Storage Conditions
Storage Temperature:
-20°C
Notes
General notes:- 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 five amino acids Asp-Asp-Asp-Asp-Lys is present just upstream of the KpnI site. This allows the protein of interest to be cleaved from maltose-binding protein with enterokinase.
- pMAL-p2E cut with KpnI followed by treatment with T4 polymerase to trim back the 3´ extension produces a blunt end at the lysine 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 enterokinase cleavage of the fusion produces a protein with no vector-derived amino acids.
- The sequences of the pMAL vectors, as well as other pMAL information, are available at www.neb.com or by e-mail from info@neb.com. A detailed map of the closely related vector pMAL-p2X can be found in the appendix of the New England Biolabs Catalog.
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.
- La Vallie, E.R. and McCoy, J.M. (1990) In Enzymatic and Chemical Cleavage of Fusion Proteins. 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. 16.4.10-16.4.11.
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-p2E 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.
This use of this product is covered by U.S. Patent No. 4,769,326 and foreign counterparts owned by the Reagents of the University of California. This product is for research use only and is not to be used for commercial purposes. Use of this product to produce products for sale or for diagnostic, therapeutic or drug discovery purposes is prohibited. In order to obtain a license to use this product for commercial purposes, contact the Regents of the University of California.
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