title: Immune response against the purified serotype specific antigen of bluetongue virus and initial attempts to clone the gene that codes for the synthesis of this protein.
abstract: Sheep were injected with different amounts of purified protein P2 of bluetongue (BT) virus (BTV). About 3 X 50 mcg was required for the induction of neutralizing antibodies. Sheep injected with 3 X 10 mcg were, however, still largely protected when challenged with virulent virus. This has suggested the possibility of using P2 as a subunit vaccine and initiated an investigation of the possibility of synthesizing P2 by DNA-recombinant technology. In order to clone the gene that codes for the synthesis of P2 both the "shotgun" approach with unfractionated dsRNA and cloning of isolated segment 2 were investigated. The basic approach was to convert the dsRNA to DNA which was cloned into the Pst 1 site of E. coli plasmid pBR322. The largest BTV-specific insert that was obtained in the initial experiments was just more than 2,000 base pairs long. The largest insert obtained when isolated segment 2 dsRNA was cloned was about 1,200 base pairs which represents about 1/3 of the P2 gene.
authors.forename: H
authors.initials: H
authors.lastname: Huismans
authors.forename: N T
authors.initials: NT
authors.lastname: van der Walt
authors.forename: B J
authors.initials: BJ
authors.lastname: Erasmus
mesh_terms: D000914:Antibodies, Viral / Q000096:biosynthesis; D000917:Antibody Formation; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D001820:Bluetongue virus / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D005814:Genes, Viral; D010957:Plasmids; D012330:RNA, Double-Stranded / Q000235:genetics; D012087:Reoviridae / Q000276:immunology*; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D004274:DNA, Recombinant; D012330:RNA, Double-Stranded; D014764:Viral Proteins