M13KE is a simple M13 derivative in which cloning siteshave been introduced at the 5´ end of gene III for display of short peptidesequences as N-terminal pIII fusions. Because this is a phage, rather than aphagemid vector, all 5 copies of pIII on the surface of each virion will befused to the cloned peptide. Since displayed proteins longer than 20–30 aminoacids have a deleterious effect on the infectivity function of pIII, this vectoris suitable only for the display of short peptides. Additionally, the vectordoes not carry a plasmid replicon or antibiotic resistance, so it is necessaryto propagate the vector as phage, rather than a plasmid (i.e., titer forplaques, not colonies). This simplifies the intermediate amplification stepsduring panning considerably, as it is not necessary to express antibiotic genesbefore plating, or to use helper phage during amplification. The steps necessaryto clone a peptide library into M13KE are outlined below. To clone a singlepeptide sequence, reactions can be scaled down.Figure 1Construction 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.
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