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Prices are in US dollars and valid only for US orders.
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Description:
The vector pMAL-c4E 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).
MBP fusions made with this vector are expressed cytoplasmically. 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 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-c4E 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.
Source:
NEB 5-alpha Competent E.coli (pMAL-c4E)
Concentration:
200 μg/ml
Storage Conditions
Storage Conditions:
10 mM Tris-HCl
1 mM EDTA
pH 7.5 @ 25°C
Storage Temperature:
-20°C
FAQs
- Why did NEB switch from selling the pMAL-p2 and c2 series of vectors to the pMAL p4 and c4 series?
References
- 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 (Eds.). Current Protocols in Molecular Biology, 16.6.1-16.6.12.
- La Vallie, E. R. and McCoy, J. M. (1990) Enzymatic and Chemical Cleavage of Fusion Proteins. In F. M. Ausebel, R. Brent, R. E. Kingston, D. D. Moore J. G. Seidman, J. A. Smith and K. Struhl (Eds.). Current Protocols in Molecular Biology, 16.4.10-16.4.11.
Legal
Patents:
New England Biolabs, Inc.: U.S. Patent No. 5,643,758
Research Use Assurance:
The buyer/user has a non-exclusive license to use the pMAL vectors for research purposes only. A license to use the pMAL vectors for commercial purposes is available from New England Biolabs, Inc.
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