FIG. 6.
Detection of in vivo gag mRNA transcription following i.m. priming and boosting with rVSV-gag vectors. Mice (n = 5 per time point) were primed by i.m. inoculation of rVSVIN vectors (1 × 107 PFU) and boosted 8 weeks later by i.m. inoculation of the corresponding GNJ protein exchange vectors. At 4 h and 1, 2, 4, and 8 days postpriming and postboosting, muscle (A) and draining lymph nodes (B) were excised from mice and homogenized in sucrose-phosphate-glutamic acid buffer. Tissue homogenates were frozen and then assessed later for Gag mRNA by quantitative real-time PCR. The limit of detection for the quantitative real-time PCR assay is 5 × 103 Gag mRNA copies per 10 mg tissue.