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Ph.D.™ Peptide Display Cloning System |
 | | | | Ph.D.™ Peptide Display Cloning System |
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Prices are in US dollars and valid only for US orders.
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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.
Kit Components:
M13 Extension Primer
M13KE Extension Primer
Storage Conditions
Storage Temperature:
-20°C
Protocols
Protocols for Ph.D.™ Peptide Display Cloning System
References
- Sambrook, J. and Russell D.W. (2001) Molecular Cloning: A Laboratory Manual, (3rd ed.), Cold Spring Harbor, Cold Spring Harbor Laboratory Press.
Companion Products
M13KE Phage
Ph.D.™-12 Phage Display Peptide Library Kit
Ph.D.™-7 Phage Display Peptide Library Kit
Ph.D.™-C7C Phage Display Peptide Library Kit
pMAL-pIII Vector
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