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Updated: Jul 10, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
Published on: March 25, 2020
In vitro selection of a 5'-purine nucleotide transferase ribozyme
1Research Center for Advanced Science and Technology, The University of Tokyo, 153-8904 Tokyo, Japan.
Abstract:
Here we report a ribozyme that catalyzes the 5'-nucleotidyl transfer reaction forming the 2'-5' phosphodiester bond. This ribozyme was retrieved as a sole sequence in the pool enriched for the 5'-triphosphate-dependent activities in incorporating ATP-gammaS. The originally selected ribozyme consisting of 109-nucleotide was miniaturized to 45-nucleotide M4 ribozyme via mutation studies, and based on this mini-ribozyme a trans-acting system was constructed. Interestingly, M4 ribozyme promiscuously accepts a variety of purine nucleotides. This remarkable ability of M4 ribozyme would lead us to the development of a new tool for the 5'-modification of RNAs with unique chemical groups.
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