Construction and analysis of ORF6-null KSHV genome.

<p>(<b>A</b>) Schematic diagrams of the structures of ORF6 and its neighboring ORFs in the wild-type and mutant BACs. (<b>B</b>) Confirmation of KSHV ORF6 replacement with Kan/SacB cassette by PCR amplification. The BAC36 (lane 1 and 3) and BACΔ6 (lane 2 and 4) were amplified with flanking PCR primers A and B and internal PCR primers C and D. (<b>C</b>) Confirmation of BAC36 and BACΔ6 genome with <i>Bg</i>lII digestion and Southern blot analysis. BAC36 (lane 1) and BACΔ6 (lane 2) DNAs were digested with <i>Bg</i>lII. The digested DNAs were electrophoresed, blotted, and hybridized with Dig-labeled ORF6 or Kan/SacB DNA probes, respectively.</p>