Journal of General Virology, Vol 79, 3145-3153, Copyright © 1998 by Society for General Microbiology

ARTICLES

Identification of epitopes in cucumber mosaic virus using a phage- displayed random peptide library

X He, S Liu and KL Perry
Department of Botany and Plant Pathology, Purdue University, West Lafayette, IN 47907, USA.

Antigenic sites in the cucumber mosaic virus (CMV) coat protein (CP) have been identified using a polyclonal antiserum prepared against glutaraldehyde-fixed virions. Antibodies were used to screen a random peptide library of heptamers displayed on the surface of a bacteriophage. Eight of 36 (22%) sequenced phage clones had inserts resembling a putative virion surface domain of the CMV CP. This region has the sequence LETDEL, corresponding to amino acids 194-199 in the Fny-CMV CP. The binding of phage clones to Fny-CMV antiserum was inhibited by a synthetic peptide representing this region. Six of 36 (17%) phage clones contained sequences corresponding to a C-terminal sequence in the Fny-CMV CP, which is thought to be internal in assembled virions. This sequence, EHQRIPTSGV, represents amino acids 206-215 and all but the P residue were observed in at least one clone. Four of 36 (11%) sequenced phage clones carried sequences that matched a portion of the sequence RLLLPDSV, corresponding to amino acids 89-96 in the Fny-CMV CP. This region was also identified as the antigenic site recognized by a monoclonal antibody (MAb23C10E4). Eleven percent of the phage (4 of 36) contained sequences matching at least three amino acids of the N-terminal region in the CMV CP. The positions of the antigenic sites seen in this study are consistent with a predicted structure for the CMV CP.

This article has been cited by other articles:

				T. J. Smith, E. Chase, T. Schmidt, and K. L. Perry The Structure of Cucumber Mosaic Virus and Comparison to Cowpea Chlorotic Mottle Virus J. Virol., August 15, 2000; 74(16): 7578 - 7586. [Abstract] [Full Text]