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  • pMAL™-c5X-His Vector

    • This product was discontinued on 01/05/2015
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    Discontinued Products

    Description

    The vector pMAL-c5X-His is designed to produce maltose-binding protein (MBP) fusions, where the protein of interest can be cleaved from MBP with the specific protease Factor Xa (NEB #P8010).

    C-terminal polyhistidine (6X-His) tag is encoded by the vector for optional addition to the target protein.

    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 maltose-binding protein). The fusion protein thus produced can be purified by amylose affinity chromatography. The sequence coding for the four amino acids Ile-Glu-Gly-Arg is present just upstream of the XmnI site. This allows the protein of interest to be cleaved from maltose-binding protein with the specific protease Factor Xa. Fragments inserted in the XmnI site (cleaves GAAGG↓ATTTC) will produce a fusion protein that, after Factor Xa cleavage, contains no vector-derived residues on the protein of interest.

    Visit DNA Sequences and Maps page for more information.


    Product Source

    NEB 10-beta Competent E. coli (pMAL-c5X-His)

    Properties and Usage

    Storage Temperature

    -20°C

    Storage Conditions

    10 mM Tris-HCl
    1 mM EDTA
    pH 7.5 @ 25°C

    Sequence Files

    Fasta  GenBank  

    Notes

    1. NEB 10-beta Competent E. coli(High Efficiency) (NEB #C3019) is recommendedfor propagation and subcloning. NEB ExpressCompetent E. coli (High Efficiency) (NEB #C2523)or NiCo21(DE3) Competent E. coli (NEB #C2529)are 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. (pp.16.6.1–16.6.12).. New York: John Wiley & Sons, Inc..
    5. Yanisch-Perron, C., Vieira, J. and Messing, J. (1985). Gene. 33, 103-119.
    6. Robichon, C., Luo, J., Causey, T.B., Benner, J.S. and Samuelson, J.C. (2011). Appl. Environ. Microbiol.. 77(13):, 4634-4646. PubMed ID: 21602383..

    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.