Figure 3.

Fluorescence detection of the PATRR-mediated rearrangement. (A) Plasmids used for the detection system. When the two plasmids harboring PATRR11 (P-SD-11) and PATRR22 (22-SA-GFP) are rearranged within each PATRR region, the transcript from the fusion product splices out the junction sequence and (middle panel) expresses the downstream GFP gene product. As a positive control, an expression vector, which includes this expected rearranged fragment, was also produced. (Arrowheads) PCR with flanking primers detected the junction fragment of ∼620 bp as early as 3 h after transfection, and (lower panel, arrow) the amount of product increased until 24 h post-transfection. (B) Flow cytometry. The GFP signal was measured 48 h after transfection. P-SD-11 and 22-SA-GFP were transfected alone or simultaneously into the cells. (Area shown in black in the histograms, R2) GFP-positive cells. (C) Percentage of GFP-positive cells. Bars, SD (n = 3).