FIG. 8.

Assembly of IN-donor complexes with target present and challenge of assembled IN-donor complexes with target. (A) IN was assembled with single-end-labeled 3.6-kbp fragment (10 ng) containing “G” sequences on the U5 end. Lanes 2 to 4, 5 to 7, and 8 to 10 contain 5, 10, and 20 nM IN, respectively. In lanes 2 to 4, IN was assembled on the donor DNA only prior to taking an aliquot for integration (Fig. 8B, top) and DNase I treatment. In lanes 5 to 7, IN was assembled with donor and then challenged with target as indicated by the arrow (50 ng per 20 μl). An aliquot was taken for integration activity (Fig. 8B, middle), and the remaining sample treated with DNase I. In lanes 8 to 10, donor, target, and IN were assembled together prior to assaying for integration activity (Fig. 8B, bottom) and DNase I treatment. Labeling is the same as in Fig. 5. (B) Strand transfer activity of samples shown in panel A.