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).