Fig 8.

trans-priming from TP and RT domain fragments by MiniRT2. The indicated TP (lanes 2 to 8) or RT (lanes 9 to 17) domain fragments (at 4 pmol each) or GST (4 pmol, as a negative control; lane 1) was added to SUMO-MiniRT2, natively purified (N, top) or refolded (R, bottom), and priming reactions were conducted in TMnNK. The trans-complementation priming reaction conducted with GST-TP (75 to 220) and GST-RT (349 to 575) served as a positive control (lane 18). The asterisks to the left of the labeled bands denote the trans-priming signals from the corresponding TP or RT domain fragments. Note that the RT domain priming signal shown in lanes 9, 10, 13, and 14 probably represented RT domain self-priming (i.e., in cis) instead of true trans-priming, as in Fig. S1A in the supplemental material (see the text for details).