Ph.D.™ Peptide Display Cloning System
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
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.
The following reagents are supplied with this product:
Store at (°C) Concentration M13KE gIII Cloning Vector 1 mg/ml M13 Extension Primer 2,150 pmol
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