Expression of human immunodeficiency virus type 1 gag gene using genetically engineered herpes simplex virus type 1 recombinants

Abstract

Infectious herpes simplex virus type 1 (HSV-1) recombinants were constructed by inserting the cDNA sequence of the human immunodeficiency virus type 1 (HIV-1) gag gene (from nucleotide position 675 [SacI] to 3859 [Asp718] of the cDNA sequences of HIV-1 strain BH-10) within the DNA sequences of the BamHI DNA fragment B of the genome of an apathogenic HSV-1 strain HFEM. This HSV-1 strain possesses a 4.1-kbp deletion within the BamHI DNA fragment B between 0.762 and 0.789 map units of the viral genome, which allows the insertion of at least 4 kbp of foreign genetic material into this particular region. The DNA sequences of the immediate early promoter (IE4) of HSV-1 that were inserted upstream from the gag gene were used as a promoter. The screening of 205 virus stocks derived from individual plaques revealed that 46 recombinant viruses harbor HIV-1 gag-specific DNA sequences. However, it was found that only six of the recombinant viruses are able to express the gag gene product of HIV-1. This indicates that the ratio of the positive recombination events is about 2.9%.