Figure 6.

Removal of 1,10-phe restores the second step of splicing. A radiolabeled Adeno transcript was incubated in nuclear extract under standard splicing conditions in the absence (lanes 1 and 2) or presence (lanes 3 and 4) of 6 mM 1,10-phe. Following 90 min at 30°C, the reactions were dialyzed against the nuclear extract buffer (Buffer D) (48) for 12 h. ATP, creatine phosphate and MgCl₂ were then added, followed by incubation for another 90 min at 30°C. RNA was purified and separated in 8% denaturing gel.