Expression of gag precursor protein and secretion of virus-like gag particles of HIV-2 from recombinant baculovirus-infected insect cells.
A recombinant baculovirus carrying the gag gene but lacking the protease coding sequences of human immunodeficiency virus type 2 (HIV-2) has been constructed. When this recombinant baculovirus is used to infect insect cells, a high level of gag precursor protein, gag pr41, is expressed. Electron microscopy showed that the majority of gag pr41 was budding through the plasma membrane and being released into the culture medium in spherical virus-like particles with a diameter of approximately 100 nm. Metabolic labeling demonstrates that gag pr41 is myristylated. Our results demonstrated that HIV-2 gag pr41 can be assembled into virus-like particles in the absence of other HIV proteins. Rabbits immunized with purified gag pr41 particles produced high-titer antibody and Western blot analysis showed that anti-gag pr41 rabbit sera recognize p17, p24, and p55 gag proteins of HIV-1. These results show that gag pr41 particles are highly immunogenic and that gag proteins of HIV-1 and HIV-2 have similar antigenic epitopes.
Luo
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L
Department of Microbiology and Immunology, University of Ottawa, Faculty of Medicine, Ontario, Canada.
Li
Y
Y
Kang
C Y
CY
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D003001:Cloning, Molecular; D015870:Gene Expression; D015683:Gene Products, gag / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors; D015498:HIV-2 / Q000235:genetics* / Q000378:metabolism; D007313:Insecta / Q000166:cytology; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009024:Morphogenesis; D014779:Virus Replication; D016335:Zinc Fingers
D015683:Gene Products, gag