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  • pMAL-p5E Vector

    Discontinued Date

    01/01/2013
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    Discontinued Products

    Description

    The vector pMAL-p5E 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 include an N-terminal signal sequence, so the fusion protein is directed to the periplasm. The MBP has been engineered for tigher 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). 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-p5E cut with KpnI followed by treatment with the Quick Blunting Kit (NEB #E1201) 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.

    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., Li, P., Riggs, P.D. and Inouye, H. (1987). Gene. 67, 21-30.
    2. 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.
    3. Nagai, K. and Thogersen, H.C. (1987). Methods Enzymology. 153, 461-481.
    4. 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.), Current Protocols in Molecular Biology. 16.6.1-16.6.12. New York: John Wiley & Sons, Inc.
    5. 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 (Ed.), Current Protocols in Molecular Biology. 16.4.10-16.4.11. New York: John Wiley & Sons, Inc.

    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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