In vitro evolution of new ribozymes with polynucleotide kinase activity.
basic_science · Level V
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Abstract
We have isolated a large number of polynucleotide kinase ribozymes from a pool of RNA molecules consisting of an ATP-binding domain flanked by regions of random sequence. Different classes of kinases catalyse the transfer of the gamma-thiophosphate of ATP-gamma S to the 5'-hydroxyl or to internal 2'-hydroxyls. An engineered version of one class is able to catalyse the transfer of thiophosphate from ATP-gamma S to the 5'-hydroxyl of an exogenous oligoribonucleotide substrate with multiple turnover, thus acting as a true enzyme.
Medical subject headings
- Biological Evolution
- Polynucleotide 5'-Hydroxyl-Kinase
- RNA, Catalytic