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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
Rift Valley fever virus, gene expression, Western blotting, antigen-antibody reactions, antigens, antiserum, diagnostic techniques, enzyme-linked immunosorbent assay, glycoproteins, glycosylation, monoclonal antibodies, nucleoproteins, protein synthesis, recombinant proteins, sheep, structural proteins, viral nonstructural proteins, virulent strains, zoonoses
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).