FIG 5.

E. chaffeensis TRP32 binds host gDNA via its tandem repeat domain. (A) TRP32 binds to host genomic DNA. Various amounts of purified host gDNA were incubated with recombinant, full-length TRP32 and then separated by EMSA. Band shifts are indicated by an arrow (lanes 5 and 6). A band shift was not observed in the corresponding lane to which TRP32 was not added (lane 9). (B) The tandem repeat domain of TRP32 is the DNA-binding domain. EMSA was performed using a TRP32 tandem repeat construct (TRP32-TR) and 5 ng of host gDNA. The TRP32 band shift is indicated by an arrow (lanes 3 and 4). Binding was abolished when excess unlabeled competitor was added (lanes 5 and 6). (C) Supershift assays in which TRP32-TR was incubated with host DNA and increasing concentrations of anti-TRP32 antibody were performed. Band shifts and super shifts are labeled with arrows.