Localization of a surface domain of the capsid protein of barley yellow dwarf virus.
We describe a method for localizing protein domains situated at the surface of virus particles. A cDNA clone of the New York PAV isolate (NY-PAV) of barley yellow dwarf virus (BYDV) containing the capsid protein gene was generated and sequenced. A defined set of overlapping cDNA fragments specific to the capsid protein ORF of NY-PAV was subcloned into the pGEX expression vectors. Cells of Escherichia coli carrying these plasmids synthesize recombinant glutathione S-transferase/capsid proteins. These proteins were used in immunoblot experiments to localize the epitopes of three PAV-BYDV-directed monoclonal antibodies to a 20 amino acid-long segment of the NY-PAV capsid protein. All three monoclonal antibodies reacted with NY-PAV virions in a triple antibody sandwich enzyme-linked immunosorbent assay, indicating that their epitopes are located at the virion surface.
Rizzo
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TM
Agricultural Research Service, U.S. Department of Agriculture, Ithaca, New York.
Gray
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SM
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D002213:Capsid / Q000235:genetics / Q000276:immunology / Q000648:ultrastructure*; D003001:Cloning, Molecular; D005814:Genes, Viral; D001467:Hordeum / Q000382:microbiology; D008969:Molecular Sequence Data; D010942:Plant Viruses / Q000235:genetics / Q000276:immunology / Q000648:ultrastructure*; D013499:Surface Properties; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D015678:Viral Structural Proteins / Q000235:genetics
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D014764:Viral Proteins; D015678:Viral Structural Proteins