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  • pMAL-c5G Vector

    Discontinued Date

    01/01/2013
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    Discontinued Products

    Description

    The vector pMAL-c5G 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).

    MBP fusions made with this vector are expressed cytoplasmically. The MBP has been engineered for tighter binding to amylose.

    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). The fusion protein produced from the vector can be purified by amylose affinity chromatography. The sequence coding for the five amino acids Pro-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-c4G 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.

    Visit DNA Sequences and Maps page for more information.

    Properties and Usage

    Affinity Tag

    Maltose-Binding Protein (MBP)

    Storage Temperature

    -20°C

    Notes

    1. NEB 10-beta Competent 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.

    References

    1. Guan, C. et al. (1987). Gene. 67, 21-30.
    2. Maina, C.V., et al. Gene. 74, 365-373.
    3. Nagai, K. and Thogersen, H.C. (1987). Methods Enzymology. 153, 461-481.
    4. Riggs, P.D. 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.), Current Protocols in Molecular Biology. 16.6.1-16.6.12. New York: John Wiley & Sons, Inc.
    5. Carter, P. et al. (1989). Proteins: Structure, Function, and Genetics. 6, 240-248.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

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