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Rift Valley Fever Virus Structural and Nonstructural Proteins: Recombinant Protein Expression and Immunoreactivity Against Antisera from Sheep

Faburay Bonto, Wilson William, McVey D. Scott, Drolet Barbara S., Weingartl Hana, Madden Daniel, Young Alan, Ma Wenjun, Richt Juergen A.
Vector borne and zoonotic diseases 2013 v.13 no.9 pp. 619-629
diagnostic techniques, nucleoproteins, zoonoses, antigens, viral nonstructural proteins, glycosylation, recombinant proteins, sheep, Western blotting, Rift Valley fever phlebovirus, gene expression, enzyme-linked immunosorbent assay, monoclonal antibodies, structural proteins, antiserum, antigen-antibody reactions, glycoproteins, virulent strains
The Rift Valley fever virus (RVFV) encodes the structural proteins nucleoprotein (N), aminoterminal glycoprotein (Gn), carboxyterminal glycoprotein (Gc), and L protein, 78-kD, and the nonstructural proteins NSm and NSs. Using the baculovirus system, we expressed the full-length coding sequence of N, NSs, NSm, Gc, and the ectodomain of the coding sequence of the Gn glycoprotein derived from the virulent strain of RVFV ZH548. Western blot analysis using anti-His antibodies and monoclonal antibodies against Gn and N confirmed expression of the recombinant proteins, and in vitro biochemical analysis showed that the two glycoproteins, Gn and Gc, were expressed in glycosylated form. Immunoreactivity profiles of the recombinant proteins in western blot and in indirect enzyme-linked immunosorbent assay against a panel of antisera obtained from vaccinated or wild type (RVFV)-challenged sheep confirmed the results obtained with anti-His antibodies and demonstrated the suitability of the baculo-expressed antigens for diagnostic assays. In addition, these recombinant proteins could be valuable for the development of diagnostic methods that differentiate infected from vaccinated animals (DIVA).