Journal
VECTOR-BORNE AND ZOONOTIC DISEASES
Volume 13, Issue 9, Pages 619-629Publisher
MARY ANN LIEBERT, INC
DOI: 10.1089/vbz.2012.1285
Keywords
Rift Valley fever virus; Structural proteins; Nonstructural proteins; Immunoreactivity profiles; Sheep
Funding
- Department of Homeland Security Center of Excellence for Emerging and Zoonotic Animal Diseases (CEEZAD) [2010-ST061-AG0001]
- Kansas Bioscience Authority
- USDA Agricultural Research Service [5430-050-005-00D]
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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).
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