FIG. 1.

Design and protein expression of rVSV-gag vectors. (A) The genomes of the rVSV vectors created for this study are diagramed in the 3′-to-5′ orientation of the negative-stranded viral RNA next to the nomenclature used for each vector. (B) Generation of rVSV vector genomic cDNAs. Endonuclease sites used for the assembly of cDNA constructs are indicated. T7-Prom represents the T7 RNA polymerase promoter sequence. Le and Tr represent the virus untranslated leader and trailer sequences, respectively. Shaded bars represent viral transcription start signals, and each viral transcription unit is separated by the nontranscribed intergenic dinucleotides GT and CT, as indicated. A T7 RNA polymerase transcription-termination signal (T7 Term) and hepatitis delta virus (HDV) ribozyme lead to the generation of a precise viral 5′ end during the virus rescue process. (C) Western blots showing in vitro HIV-1 Gag expression of rVSV vectors 24 h postinfection of replicate BHK cell monolayers. Lanes 1 and 2 contain proteins from uninfected and “empty” rVSV-infected cells, respectively. Lanes 3 to 7 contain proteins from cells infected with the different Gag-expressing rVSV vectors. Protein size markers (24 kDa to 50 kDa) were run alongside test samples.