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M94A2064.TXT
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1994-10-24
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Document 2064
DOCN M94A2064
TI Incorporation of gp160 in SIVmac virus-like particles assembled from gag
in insect cells and larvae.
DT 9412
AU Ahmad S; Giavedoni L; Gardner M; Luciw P; Yilma T; Uni. of California,
Davis 95616.
SO Int Conf AIDS. 1994 Aug 7-12;10(1):39 (abstract no. 130A). Unique
Identifier : AIDSLINE ICA10/94370511
AB OBJECTIVE: We inquire incorporation of surface glycoprotein (gp160) into
the envelope of VLPs during the co-expression of gp160 and gag of
SIVmac239 in insect cells (Sf9) and larvae. Large quantities of these
VLPs are produced for use in vaccine trials. METHODS: The gp160 and gag
genes were inserted separately or together into the genome of Autographa
californica nuclear polyhedrosis virus (AcNPV) under polyhedrin promoter
by homologous recombination. AcNPV expressing gp160 or gag and AcNPV
expressing gp160 and gag were used to infect the insect cells and larvae
for characterization of recombinant gp160, gag, and VLPs from Sf9 cells
by biochemical and immunological methods. RESULTS: Electron microscopic
(EM) examination of the infected Sf9 cells showed the assembly and
release of VLPs from the plasma membrane surfaces. The expression of
VLPs by baculovirus is so high that there is particle formation on the
membranes even inside the cytoplasm. Milligram quantities of these VLPs
were produced in insect cell culture. The western-blot analysis of
sucrose gradient-purified VLPs contained both gag and env proteins. EM
studies with anti-gp160 immunogold labeling of purified VLPs are in
progress. DISCUSSION AND CONCLUSIONS: We demonstrate the incorporation
of gp160 into envelope of VLPs assembled from gag. The incorporation of
gp160 into the envelope of VLPs does not require any SIV factor.
DE Animal Cells, Cultured *Genes, gag Insects/CYTOLOGY
Larva/MICROBIOLOGY Nuclear Polyhedrosis Virus/*GENETICS Promoter
Regions (Genetics) SIV/*GENETICS Viral Envelope Proteins/*ISOLATION &
PURIF Virion/*GENETICS MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).