Design and optimization of effector-activated ribozyme ligases

摘要: A selected ribozyme ligase, L1, has been engineered to respond small organic effectors. Residues important for catalysis were mapped a compact core structure. Aptamers that bound adenosine and theophylline appended the structure, resultant aptazymes proved be responsive their cognate Rational sequence substitutions in joining region between aptamer yielded whose activities enhanced from 800–1600-fold presence of 1 mM ATP or theophylline, respectively. However, when an anti-flavin was structure flavin-responsivity minimal. The randomized series negative positive selection steps activated by up 260-fold 100 µM FMN. regions ‘communication modules’ could used join other aptamers form aptazymes. These results show ligases can readily function as allosteric enzymes, reveal many techniques principles previously demonstrated during development hammerhead may generalizable.