Expression of bluetongue virus group-specific antigen VP3 in insect cells by a baculovirus vector: its use for the detection of bluetongue virus antibodies.
DNA representing RNA segment 3 of bluetongue virus (BTV) serotype 17, corresponding to the gene that codes for a group-specific antigen VP3, has been inserted into a baculovirus transfer vector in lieu of the 5' coding region of the polyhedrin gene of Autographa californica nuclear polyhedrosis virus (AcNPV). After cotransfection of Spodoptera frugiperda cells with wild-type AcNPV DNA in the presence of the derived recombinant transfer vector DNA, polyhedrin-negative recombinant baculoviruses were recovered. When S. frugiperda cells were infected with one of these recombinant viruses, a protein that was similar in size and antigenic properties to the BTV VP3 protein was synthesized. Antibodies raised in mice or rabbits to the baculovirus-expressed VP3 protein immunoprecipitated the VP3 protein of BTV-17 as well as that of BTV-10. The expressed antigen reacted with antisera representing four U.S.A. BTV serotypes in an indirect ELISA test.
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D001820:Bluetongue virus / Q000276:immunology*; D002460:Cell Line; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005814:Genes, Viral; D005822:Genetic Vectors; D007304:Insect Viruses / Q000235:genetics*; D009036:Moths; D009693:Nucleic Acid Hybridization; D012087:Reoviridae / Q000276:immunology*; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015678:Viral Structural Proteins
D000914:Antibodies, Viral; D000956:Antigens, Viral; D004279:DNA, Viral; D014764:Viral Proteins; D015678:Viral Structural Proteins; D004262:DNA Restriction Enzymes