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Journal of Virology, October 1999, p. 8496-8502, Vol. 73, No. 10
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

The Capsid Gene of Feline Calicivirus Contains Linear B-Cell Epitopes in both Variable and Conserved Regions

Alan D. Radford,1,* Kim Willoughby,1 Susan Dawson,2 Christina McCracken,1 and Rosalind M. Gaskell1

Department of Veterinary Pathology1 and Department of Veterinary Clinical Sciences and Animal Husbandry,2 University of Liverpool, Veterinary Teaching Hospital, Leahurst, Neston CH64 7TE, United Kingdom

Received 24 May 1999/Accepted 12 July 1999

In order to map linear B-cell (LBC) epitopes in the major capsid protein of feline calicivirus (FCV), an expression library containing random, short (100- to 200-bp) fragments of the FCV F9 capsid gene was constructed. Analysis of this library showed it to be representative of the region of the capsid gene that encodes the mature capsid protein. The library was screened by using polyclonal antisera from a cat that had been challenged experimentally with F9 to identify immunoreactive clones containing LBC epitopes. Twenty-six clones that reacted positively to feline antisera in immunoblots were identified. FCV-derived sequence from these clones mapped to a region of the capsid that spanned 126 amino acids and included variable regions C and E. An overlapping set of biotinylated peptides corresponding to this region was used to further map LBC epitopes by using F9 antisera. Four principal regions of reactivity were identified. Two fell within the hypervariable region at the 5' end of region E (amino acids [aa] 445 to 451 [antigenic site {ags} 2] and aa 451 to 457 [ags 3]). However, the other two were in conserved regions (aa 415 to 421 [ags 1; region D] and aa 475 to 479 [ags 4; central region E]). The reactivity of the peptide set with antisera from 11 other cats infected with a range of FCV isolates was also determined. Ten of 11 antisera reacted to conserved ags 4, suggesting that this region may be useful for future recombinant vaccine design.

* Corresponding author. Mailing address: Department of Veterinary Pathology, University of Liverpool, Veterinary Teaching Hospital, Leahurst, Neston CH64 7TE, United Kingdom. Phone: 44 151 794 6017 or 6012. Fax: 44 151 794 6005. E-mail: alanrad{at}liv.ac.uk.

Journal of Virology, October 1999, p. 8496-8502, Vol. 73, No. 10
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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