Expression of Foreign Proteins on the Surface of Autographa Californica Nuclear Polyhedrosis Virus. (April 1997)

• Record Type: Journal Article
• Authors: Grabherr, R.; Ernst, W.; Doblhoff-Dier, O.; Sara, M.; Katinger, H.
• Is Part Of: Biotechniques. Volume 22:Number 4(1997)
• Page Start: 730
• Page End: 735
• Publication Date: 1997-04
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/97224rr02
• Languages: English
• ISSNs: 0736-6205

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.