(A) Analysis of the gD-encoding gene in HSV SC16 gD-YFP. DNAs from SC16 gD-YFP and wild-type (wt) SC16 were analyzed by PCR with primers within the coding sequence of gD and near the start of the gene for gI. Products were separated on a 0.8% agarose gel. Lanes: 1, 1-kb molecular size marker; 2, HSV-1 SC16; 3, HSV-1 SC16 gD-YFP. (B) Western blot analysis of cell lysates infected with wild-type SC16 or SC16 gD-YFP separated on a 12% polyacrylamide gel, transferred to nitrocellulose membranes, and probed with either anti-gD LP2 or an anti-GFP MAb which reacts with YFP. (C) Western blot analysis of virions pelleted (100,000 × g) from wild-type SC16- or SC16 gD-YFP-infected cell supernatants separated on a 12% polyacrylamide gel, transferred to a nitrocellulose membrane, and probed with anti-gD MAb LP14.