Induction of antibodies to the Epstein-Barr virus glycoprotein gp85 with a synthetic peptide corresponding to a sequence in the BXLF2 open reading frame.
Epstein-Barr virus codes for at least three envelope glycoproteins, one of which, gp85, has not yet been mapped to the viral genome. The publication and analysis of the entire Epstein-Barr virus DNA sequence has allowed identification of open reading frames with potential for encoding membrane glycoproteins. To determine whether one of these candidate open reading frames, BXLF2, codes for gp85, an antibody was made to a 17-residue peptide derived from positions 518 to 533 of the predicted BXLF2 protein. The reactivity of the antipeptide antibody was then compared with that of the monoclonal antibody F-2-1, which was originally used to define and characterize gp85. Antipeptide antibody and F-2-1 immunoprecipitated glycosylated molecules with identical electrophoretic mobilities; digestion of the two immunoprecipitated proteins with V8 protease generated comparable peptides; and the antipeptide antibody reacted in Western immunoblots with the gp85 glycoprotein that had been immunoprecipitated by F-2-1 before transfer to nitrocellulose. In addition, a monospecific rabbit antibody, made against native gp85, reacted with the peptide used for immunization. These results are compatible with the hypothesis that the BXLF2 open reading frame codes for gp85.
Oba
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Department of Comparative and Experimental Pathology, University of Florida, Gainesville 32610.
Hutt-Fletcher
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LM
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000276:immunology; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005814:Genes, Viral; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology*; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007118:Immunoassay; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D006023:Glycoproteins; D010455:Peptides; D014759:Viral Envelope Proteins