Figure 7

PCR Analysis of HSV-2 BAC recombinants. (A) Insert-specific PCR of 4 individual clones each of HSV2-BAC recombinants containing an inserted DsRed2 (left panel), vaccinia virus B5R (middle panel) or vaccinia virus A33R gene (right panel). Lane M -100 bp molecular size markers. Lane P – Specific plasmid controls containing the genes for DsRed2, B5R, A33R, respectively (from left to right). Sizes of PCR products are: DsRed2 – 424 bp; B5R – 1014 bp; A33R – 586 bp. (B) UL41 and EGFP PCR analysis of 4 individual clones each of HSV2-BAC recombinants containing an inserted vaccinia virus B5R (lanes 1–4), vaccinia virus A33R gene (lanes 5–7), or DsRed2 gene (lanes 9–12). Lane M -100 bp molecular size markers. Lane 13 – HSV2-BAC. Lane 14 – DEST-BAC. The top half of the gel shows the PCR products generated with UL41 primers for each clone. Sizes of UL41 PCR products are: B5R – 2280 bp; A33R – 1850 bp; DsRed2 – 1290 bp (DsRed2); HSV2-BAC – 342 bp; UL41 PCR of DEST-BAC – 1320 bp). The bottom half of the gel shows the PCR products using EGFP primers for each clone, confirming the absence of the EGFP in each recombinant. Size of EGFP PCR product of DEST-BAC is 433 bp (lane 14).