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  • pMAL-c5E Vector

    • This product was discontinued on 01/05/2015
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    Description

    The vector pMAL-c5E 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). 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-c5E 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.

    DNASU is a central repository for plasmid clones and collections that may also be helpful.

    Properties and Usage

    Affinity Tag

    Maltose-Binding Protein (MBP)

    Storage Temperature

    -20°C

    Storage Conditions

    10 mM Tris-HCl
    1 mM EDTA
    pH 8.0 @ -20°C

    Sequence Files

    Fasta  GenBank  

    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.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

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    Safety Data Sheet

    The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.