Analysis of glycoproteins of viruses in the family Bunyaviridae.

The membrane glycoproteins (Gn and Gc) of viruses in the family Bunyaviridae form projections on the virion envelope and are involved in virus entry and eliciting protective immunity. The glycoproteins are modified by N-linked glycosylation and accumulate in the Golgi complex where virions mature and bud. In this chapter, we describe the methods that have been used in our laboratory for the study of the glycoproteins of Bunyamwera virus, the prototype of the family. The protocols cover the expression of viral glycoproteins, examination of intracellular localization by immnunofluorescent confocal microscopy, radiolabeling, immunoprecipitation, and SDS-PAGE analysis of the proteins, and the improved reverse genetic system to rescue recombinant viruses that contain mutations at N-linked glycosylation sites.