FIG. 3.

Southern blot characterization of the recombinant viruses. Viral DNAs from KOS (lane 1), KgBpK⁻ (lane 2), KCZ (lane 3), and KgBpK⁻gC⁻ (lane 4), as well as viral DNAs from KgBpK⁻gC^R and KgBpK^RgC⁻, were digested with the restriction endonuclease BamHI (A) or NcoI (B and C) and subjected to Southern blot analysis. (A) A ³²P-labeled gB probe hybridized to a 7,774-bp fragment of wild-type gB sequence encoded by KOS (lane 1) and KCZ (lane 3), and KgBpK^RgC⁻ (lane 6) hybridized with a smaller fragment of 3,009 bp in the recombinant viruses KgBpK⁻ (lane 2), KgBpK⁻gC⁻ (lane 4), and KgBpK⁻gC^R (lane 5) due to the introduction of a BamHI restriction endonuclease site within the gB sequence at the site of the deletion of amino acids 68 to 76. (B) A ³²P-labeled gC probe (642-bp NcoI fragment of pgC1) encoding the deleted gC sequence hybridized to a 642-bp fragment containing wild-type gC in KOS (lane 7), KgBpK⁻ (lane 8), and KgBpK⁻gC^R (lane 11). (C) A different gC probe (828-bp NcoI fragment of pgC1 undeleted in all viruses) hybridized with an 11.2-kbp fragment in KCZ (lane 15), KgBpK⁻gC⁻ (lane 16), and KgBpK^RgC⁻ (lane 18), in which the gC coding sequence was deleted and replaced with the human cytomegalovirus immediate early promoter driving the lacZ gene, and it hybridized with an 828-bp fragment in KOS (lane 13), KgBpK⁻ (lane 14), and KgBpK⁻gC^R (lane 17), in which the wild-type gC coding sequence was present.