Figure 1.

Construction of rHSVrep/cap. (A) The cloning strategy into the single XbaI site at nucleotide position 143.969 of HSV-1, strain 1802 flanked by non-essential HSV genes is depicted. The AAV-2 rep and cap gene expression cassette (nucleotides 191–4485) includes the AAV promoters and polyA⁺ signal, and is linked to an SV40 controlled lacZ gene. LacZ positive HSV clones were plaque purified and analyzed for AAV Rep and Cap expression. Lysates of rHSVrep/cap-infected cells were prepared at different times p.i. Samples equaling 1 × 10⁴ cells were loaded per lane. Western blots were reacted with the indicated mAb, followed by peroxidase-coupled second antibody and enhanced chemiluminescence. (B) AAV capsid (VP) proteins analyzed with mAb B1. (C) AAV Rep detected with mAb 303.9. HSV + AAV infected cell extracts were prepared at 12 h p.i.