FIG 8.

Binding of K8 to ori-Lyt DNA is mediated by T1.4 RNA. (A) Schematic diagrams of the T1.4 locus in the KSHV genome and strategy for construction of BAC-ΔT1.4. The nucleotide sequences refer to GenBank accession number U75698.1. The sequence chromatogram of the deletion junction in BAC-ΔT1.4 is shown. (B) Electrophoretic analysis of the BAC-ΔT1.4 mutant viral genomes. BAC16 (BAC WT) and BAC-ΔT1.4 DNAs were digested with XhoI, resolved on a 0.8% agarose gel, and stained with ethidium bromide. The red arrow indicates the changes of restriction pattern caused by the deletion of T1.4 in BAC-ΔT1.4. (C) BAC16 and BAC-ΔT1.4 were introduced into 293T cells, and the cells were induced with sodium butyrate for 48 h. A ChIP assay was performed with anti-K8 and anti-RTA antibodies or IgG and detected by PCR with primers for the 12F region or GAPDH. The data are shown as the means ± SD of the results of three separate experiments. *, P < 0.05.