Characterization of Dugbe virus by biochemical and immunochemical procedures using monoclonal antibodies
Dugbe virus is assigned to the family Bunyaviridae, genus Nairovirus and is related to the tick-borne Crimean-Congo hemorrhagic fever and Nairobi sheep disease viruses. The proteins of Dugbe virus were studied biochemically and immunochemically using monoclonal and poly-clonal antibodies. The G1, N and G2 proteins were detected by PAGE at 73, 49, and 35 kDa, respectively. On a Western blot, polyclonal antisera to Dugbe virus reacted with the G1, N and G2 proteins and with several additional proteins (210, 45, 40, 33, and 30 kDa). The Gl and G2 proteins were shown to be located on the surface of virus particles and to be glycosylated while the N protein was internal and non-glycosylated. The 40 kDa protein also was found to be glycosylated. This 40 kDa glycoprotein may represent an additional glycoprotein, as found in Hazara virus, or may be a breakdown product of G1. A monoclonal antibody (McAb H28.89) against the N protein specifically labelled purified nucleocapsids in an immunogold EM reaction. This McAb did not neutralise viral infectivity in an in vitro assay and did not label whole virus particles with colloidal gold. Another McAb (H28.17) against the G1 protein neutralised virus infectivity and labelled whole virus particles by immunogold EM.
This word, combined with ongoing genome sequencing and expression research, should lead to a better understanding of the molecular biology of nairoviruses and to the production of useful diagnostic tools.
KeywordsVirus Particle Colloidal Gold Polyclonal Antiserum Nucleocapsid Protein Intact Virus Particle
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