FIG. 7.

Effects of mutations of the C/EBP motifs in the ori-Lyt fragment (3F) on recruiting of replication proteins to ori-Lyt DNA. (A) Schematic diagram of the sequences and locations of eight C/EBP binding sites that are organized as four spaced palindromes, as well as site-specific mutations on these C/EBP motifs. (B) Biotinylated DNA fragments were prepared by PCR with a pOri-A (wild-type) DNA template as well as pOri-M12 and pOri-M1256, mutations on certain C/EBP motifs, which were described by Wang et al. (31). TPA-induced BCBL-1 nuclear extract (NE) was incubated with the DNA fragments conjugated on magnetic beads, washed, and eluted with D500 elution buffer. Samples were assayed by Western blotting with antibodies as indicated. An irrelevant DNA was included as a control, C.