Production of human immunodeficiency virus (HIV)-like particles from cells infected with recombinant vaccinia viruses carrying the gag gene of HIV.
We constructed a recombinant vaccinia virus carrying the entire gag and pol genes of human immunodeficiency virus type 1 (HIV-1). The main gene product detected in the lysates of infected CV-1 and SW480 cells was the gag precursor protein. However, in the culture fluid of infected SW480 cells, but not of infected CV-1 cells, reverse transcriptase (RT) activity was detected. The highest RT activity was found at a density of 1.15 g/ml and this fraction contained many round particles with diameters of 100-150 nm. In contrast to the infected cell lysates, the particles contained the processed gag and pol proteins, suggesting that particle formation may be a prerequisite for efficient processing of the gag precursor by the HIV protease encoded in the pol gene. Particles were also recovered from the culture fluid of SW480 cells infected with another recombinant vaccinia virus carrying only the gag gene. These particles contained the unprocessed gag precursor, indicating that the gag precursor alone was sufficient for particle production.
Shioda
T
T
Department of Viral Infection, University of Tokyo, Japan.
Shibuta
H
H
D000818:Animals; D002460:Cell Line; D004251:DNA Transposable Elements; D004279:DNA, Viral / Q000235:genetics; D015685:Fusion Proteins, gag-pol / Q000235:genetics; D015683:Gene Products, gag / Q000235:genetics*; D005814:Genes, Viral; D015751:Genes, pol; D015497:HIV-1 / Q000235:genetics* / Q000302:isolation & purification; D007700:Kinetics; D010957:Plasmids; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D011995:Recombination, Genetic; D014616:Vaccinia virus / Q000235:genetics*; D015678:Viral Structural Proteins / Q000235:genetics*
D004251:DNA Transposable Elements; D004279:DNA, Viral; D015685:Fusion Proteins, gag-pol; D015683:Gene Products, gag; D015678:Viral Structural Proteins; D012194:RNA-Directed DNA Polymerase