Table 1.
Optimized TES reaction conditions, product yields and reaction rates
| Optimized TES reactions conditionsa |
TES product yield and observed rate constants for substrate cleavage and exon ligation reactionsb |
||||||
|---|---|---|---|---|---|---|---|
| Ribozyme | MgCl2 (mM) | Time (min) | Ribozyme (nM) | TES product (%) | Substrate cleavage (kobs/min) | Exon ligation (kobs/min) | |
| Pneumocystis | 15 | 30 | 100 | 68.5 ± 4.4 | 5.0 ± 1.1 | 3.2 ± 0.1 | |
| Tetrahymena | 10 | 45 | 166 | 47.5 ± 0.18 | 0.17 ± 0.05 | 0.14 ± 0.02 | |
| Candida | 25 | 30 | 75 | 50.0 ± 1.5 | 0.31 ± 0.01 | 0.34 ± 0.03 | |
aTES reactions were optimized with regard to magnesium concentration, time and ribozyme concentration for maximum TES product yields.
bThe observed rate constants, kobs, were obtained for the substrate cleavage and exon-ligation reactions under single turnover conditions. Note that ribozyme and substrate sequences are different between the ribozyme constructs.