Figure 3
From: Ligand-induced sequestering of branchpoint sequence allows conditional control of splicing
Insertion of BPS in theophylline-binding aptamer does not activate cryptic branchpoint. Lariat intermediate isolated from preparative splicing reactions of AdBPT15AG pre-mRNA performed in the absence (lanes 6 and 7) or presence of theophylline (lanes 8 and 9) was subjected to branchpoint mapping. Primer extension was performed on intact (lane 6, -S100) or debranched lariat intermediate (lanes 7 and 9, +S100). Asterisks (*) represent primer extension products from S100 total RNA (lane 10). The primer extension products were separated on a 13% denaturing polyacrylamide gel. The dideoxy-sequencing ladder from parental DNA is shown in lanes 2–5. M, 10 bp DNA ladder (Invitrogen).