FIG. 6.

Synthesis of full-site DNA products using a 3.6-kbp donor possessing wt U3 and mutant U5 att ends. (A) The modified U5 att end on the 3.6-kbp donor is described in Fig. 7, row B. Standard reaction conditions were used as described in the legend to Fig. 3 using IN at 5 nM. The graph depicts the amount of full-site and half-site products produced versus time of incubation at 37°C. (B) The U5 end of the 3.6-kbp DNA donor was modified as described in Materials and Methods and as shown in Fig. 7. HIV-1 IN (5 nM) was assembled with labeled donor DNA for 15 min at 14°C. Target DNA was added for strand transfer at 37°C, and the products were removed and processed after 120 min. The products were and were not subjected to EcoRI digestion (top). Lanes 1 and 2 were mutant DNA without IN, while lanes 3 and 4 were with IN. The half-site and full-site products (bold) in lane 4 produced by EcoRI digestion are identified on the top right and bottom right, respectively. The positions of the 1-kbp DNA ladder marker (not bold) are also illustrated on the right.