Expression of foreign proteins on the surface of Autographa californica nuclear polyhedrosis virus.
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 of baculovirus particles. We chose the glycosylated envelope protein gp41 of human immunodeficiency virus type 1 (HIV-1) as a model for displaying recombinant proteins on budded virus. The ectodomain of the envelope protein gp41 of HIV-1 was being fused to the entire baculovirus major coat protein gp64 (Ac-cops41) and to the membrane anchor sequence of gp64 (Acmars41). 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.
Grabherr
R
R
University of Agriculture, Food Science and Forestry, Vienna, Austria.
Ernst
W
W
Doblhoff-Dier
O
O
Sara
M
M
Katinger
H
H
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D005822:Genetic Vectors; D015700:HIV Envelope Protein gp41 / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D008854:Microscopy, Electron; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics*; D018411:Spodoptera / Q000235:genetics; D014760:Viral Fusion Proteins / Q000235:genetics*
D015700:HIV Envelope Protein gp41; D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins