Identiˆcation by the Phage-display Technique of Peptides That Bind to H7 Flagellin of Escherichia coli
نویسندگان
چکیده
The four peptides interacting with H7 ‰agellin of Escherichia coli were selected from a phage display library. The library was selected four times, and the interacting phage peptides were competitively eluted with H7 ‰agellin. An enzyme-linked immunosorbent assay (ELISA) showed that these peptides were reactive with the H7 ‰agellin in a dose-dependent manner. Among them, a D1 phage clone showed the highest binding a‹nity to the H7 ‰agellin. We synthesized the D1 peptide (LHIHRPTLSIQG) corresponding to the peptide-encoding region of the D1 phage clone. The synthetic peptide showed micro-molar a‹nity (EC50 value=1.9 mM) for the H7 ‰agellin. Furthermore, this D1 peptide interacted more speciˆcally with the H7 ‰agellin than with the other ‰agellins (H1, H5, H12, or H23) of E. coli. In situ hybridization clearly showed that the peptide only detected those cells harboring the H7 ‰agellin gene ( ‰iC). The peptide may speciˆcally bind to the H7 ‰agellin on the cell surface. These results suggest that the phage-display technique could be used as a tool for identifying peptides as an alternative to using a ligand as a diagnostic reagent in food products or in clinical testing.
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