The Ph.D.™ Peptide Display Cloning System contains M13KE vector, the parent vector of NEB’s Ph.D. libraries. The included M13 extension primer is used to make DNA duplex out of a user provided synthetic oligo, with or without random bases. The display system is best used to display small peptides, i.e. sequences of 50 amino acids or less.
M13KE Vector is based on M13mp18 containing the entire M13 genome, with specific cloning sites at the 5’ gIII, encoding pIII coat protein
Allows for user-designed custom peptide library construction
May be transfected into F’ E. coli to obtain infectious phage
Product Information
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.
This product is related to the following categories:
Kaur K, Taneja NK, Dhingra S, Tyagi JS (2014) DevR (DosR) mimetic peptides impair transcriptional regulation and survival of Mycobacterium tuberculosis under hypoxia by inhibiting the autokinase activity of DevS sensor kinase BMC Microbiol; 14, 195. PubMedID: 25048654, DOI: 10.1186/1471-2180-14-195
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New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
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