• My NEB
  • Print
  • PDF
  • Ph.D.™ Peptide Display Cloning System

    Description

    M13KE is a simple M13 derivative in which cloning sites have been introduced at the 5´ end of gene III for display of short peptide sequences as N-terminal pIII fusions. Because this is a phage, rather than a phagemid vector, all 5 copies of pIII on the surface of each virion will be fused to the cloned peptide. Since displayed proteins longer than 20–30 amino acids have a deleterious effect on the infectivity function of pIII, this vector is suitable only for the display of short peptides. Additionally, the vector does not carry a plasmid replicon or antibiotic resistance, so it is necessary to propagate the vector as phage, rather than a plasmid (i.e., titer for plaques, not colonies). This simplifies the intermediate amplification steps during panning considerably, as it is not necessary to express antibiotic genes before plating, or to use helper phage during amplification. The steps necessary to clone a peptide library into M13KE are outlined below. To clone a single peptide sequence, reactions can be scaled down.

    Figure 1.
    Construction of a peptide library in M13KE. Schematic shows the sequence of the peptide cloning site as well as the strategy for designing and cloning a peptide library into M13KE. The sequence of the extension primer is outlined. N = A, G, C or T; X = any user defined or randomized amino acid.

    System Components

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    M13 Extension Primer2,150 pmol
    M13KE gIII Cloning Vector

    References

    1. Sambrook, J. and Russell D.W (2001). Molecular Cloning: A Laboratory Manual, (3rd ed.). Cold Spring Harbor, Cold Spring Harbor Laboratory Press.

    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.

    Manuals

    The Product Manual 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 these document files: manual[Catalog Number].

    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.
    1. Can cDNA libraries be cloned into M13KE?
    2. Can I display antibody fragments or proteins at the N-terminus of pIII coat protein with M13KE? 
    3. What are the differences between pIII and pVIII display?
    4. Does the Ph.D.™ Phage Display Cloning System come with double-stranded DNA vector?
    5. What type of strain do I use with the Ph.D.™ Phage Display Cloning System?
    6. I have done all the standard troubleshooting for my cloning and transfection steps and I still cannot get the desired clone in M13KE?
    1. Protocol for Cloning Peptide Display Libraries in M13KE
    2. M13 Amplification

    Troubleshooting Guides

    Application Notes