Characterization of glycoproteins from insect specific Goutanap and Negev viruses

Negevirus is a recently discovered insect specific virus taxon. We focus on characterization of the viral proteins, structural organization of the virion and the principles of their interaction with the host cell. ORF2 and ORF3 (1.2 kbp and 0.7 kbp long) are predicted to encode membrane glycoproteins. We have cloned and overexpressed ORF2 and ORF3 of Goutanap and Negev viruses using inducible insect expression system. Furthermore, we have defined, successfully produced and purified recombinant soluble ectodomains of ORF2 for both viruses using affinity chromatography. Subsequent characterization using gel filtration, ion exchange and MALS techniques reveal that ORF2 ectodomains of Goutanap and Negev viruses exhibit different higher order assembly patterns: dimerization, and multimerization, in a concentration and pH dependent manner that presumably correspond to their biological role.