Expression of Foreign Proteins on the Surface of Autographa Californica Nuclear Polyhedrosis Virus

Abstract

Based on the method of direct cloning into the baculovirus genome by linearizing and re-ligation in presence of the target insert, we designed viral constructs that express foreign genes on the surface o baculovirus particles. We chose the glycosylated envelope protein gp41 of human immunodeficiency virus type 1 (HIV-1) as a model for displaying recombinant protein on budded virus. The ectodomain of the envelope protein gp41 of HIV-1 was being fused to the entire baculovirus major coa protein gp64 (Ac-cops41) and to the membrane anchor sequence of gp64 (Ac-mars41). Two different promoters, the “very late” polyhedrin promoter (Ac-mars41) and the “early and late” gp64 promoter (Ac-promars41) were compared. The expression of gp41 in infected cells and its presence on viral particles was confirmed by enzyme-linked immunosorbent assay (ELISA), Western blot and electron microscopy.