title: Selection of phage-display peptides that bind to cucumber mosaic virus coat protein.
abstract: Several discrete peptides that bind specifically to the coat protein of cucumber mosaic virus (CMV) were isolated from a diverse phage library displaying random nonapeptides on the major coat protein VIII. Enrichment was shown by polyclonal phage enzyme linked immunosorbent assay (ELISA) after three rounds of selection. Sequencing of the genes encoding 10 of these peptides revealed an absence of any conserved motifs, although nine of them contained a high proportion of proline residues. Some of the selected peptides were displayed at the N-terminus of thioredoxin and expressed in the cytoplasm of Escherichia coli. Both the phage-displayed and thioredoxin-fusion versions of the peptides could detect purified CMV and CMV present in crude leaf extracts from infected plants. By dot blot analysis, a thioredoxin-peptide fusion could readily detect as little as 5 ng of CMV. The peptides did not bind to other plant viruses. These peptides have been shown to be specific and highly sensitive tools in the detection of CMV and, as well as their diagnostic potential, they could form the basis for a novel disease resistance strategy.
authors.affiliation: Department of Biology, University of Leicester, Leicestershire, UK. kcg3@le.ac.uk
authors.forename: K C
authors.initials: KC
authors.lastname: Gough
authors.forename: W
authors.initials: W
authors.lastname: Cockburn
authors.forename: G C
authors.initials: GC
authors.lastname: Whitelam
mesh_terms: D000595:Amino Acid Sequence; D019677:Artificial Gene Fusion; D001435:Bacteriophages / Q000235:genetics; D002213:Capsid / Q000235:genetics / Q000378:metabolism*; D036022:Capsid Proteins; D017799:Cucumovirus; D015870:Gene Expression; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D013879:Thioredoxins / Q000235:genetics
chemical_list: D036022:Capsid Proteins; D019151:Peptide Library; D010455:Peptides; D011993:Recombinant Fusion Proteins; C119217:coat protein pVIII, Enterobacteria phage fd; D013879:Thioredoxins