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  • pMAL-pIII Vector

    Discontinued Date

    01/05/2015
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    Categories:
    Discontinued Products

    Description

    The pMAL-pIII Vector is a derivative of pMAL-p2 in which the leader sequence of maltose binding protein (MBP, malE) has been replaced with the M13 pIII leader sequence. KpnI/Acc65I and EagI sites have been introduced within the pIII leader to facilitate direct transfer of sequences selected from any of the Ph.D. phage display peptide libraries into an expression vector. The corresponding peptides are expressed as N-terminal MBP fusions which bear some similarity to M13 pIII displayed peptides.These fusions can then be easily purified from E. coli periplasmic space by osmotic shock followed by affinity chromatography on amylose resin. Since the peptide is expressed at the N-terminus of MBP, Factor Xa cannot be used to cleave the peptide from MBP.
     
    N8101
    pMAL-pIII cloning region. Note PhD fragments are inserted into the M13KE pIII leader sequence, which is directly followed by the malE gene.

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

    Advantages and Features

    Features

    • Alternative to chemical peptide synthesis
    • Subcloning of Ph.D peptides into pMAL-pIII provides monovalent MBP fusion for additional binding studies after phage panning, phage ELISA experiments.

    Properties and Usage

    Affinity Tag

    Maltose-Binding Protein (MBP)

    Storage Temperature

    -20°C

    Sequence Files

    Fasta  GenBank  

    Notes

    1. NEB Express Competent E.coli (High Efficiency) (NEB #C2523) is recommended for expression using this vector or TB1 (NEB #E4122).

    References

    1. Zwick, M. B. et al (1998). Anal Biochem. 264, 87-97.
    2. Zagursky, R.J. et al (1984). Gene. 27, 193-91.

    FAQs

    1. What would be a useful sequencing primer to confirm presence of my insert at the 5’ end of the malE gene?

    Protocols

    1. pMal-pIII Vector Protocol

    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.

    Application Notes

    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.