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Mol Biol (Mosk) ; 24(6): 1666-74, 1990.
Article in Russian | MEDLINE | ID: mdl-2094814

ABSTRACT

Monkey kidney cells CV-1 were infected with recombinant vaccinia virus carrying HIV-1 gag gene with a deletion of 230 nucleotide pairs from the 3'-terminus. The main gene product detected in the lysates of infected cells was the gag precursor rp50. The protein was accumulated on the cell membranes suggesting that it had a myristylated N-terminus, and was cleaved by a recombinant virus specific protease with the formation of two proteins, p17 and p24 corresponding in molecular masses to mature gag proteins. Virus-like particles similar to immature HIV virions were budding from the surface of infected cells. They look like the ring of optically dense material covered with a lipid bilayer, of the same size (100-120 nm) and of the same density in a sucrose gradient (1.16-1.18 g/ml) as HIV-1 virions. The particles contained rp50 and cellular heterogeneous RNA. Thus, the unprocessed gag precursor with deleted 77 amino acid residues from the C-terminus is able to form virus-like particles in the absence of env proteins and virus-specific RNA, and these particles are budding from the cell surface. The question about the use of extracellular Gag-particles for AIDS diagnostic work and construction of vaccines is discussed.


Subject(s)
Gene Expression Regulation, Viral , Gene Products, gag/genetics , HIV-1/genetics , Vaccinia virus/genetics , Virion , Blotting, Northern , Cell Line , Electrophoresis, Polyacrylamide Gel , Hydrolysis , Microscopy, Electron , Microscopy, Fluorescence , Nucleic Acid Hybridization , RNA, Viral/analysis , Recombination, Genetic
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