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Updated: Jul 11, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
Published on: March 25, 2020
A simple selection strategy for evolving highly efficient enzymes
Martin Neuenschwander1, Maren Butz, Caroline Heintz
1Laboratory of Organic Chemistry, ETH Zurich, Hönggerberg HCI F 339, CH-8093 Zurich, Switzerland.
Abstract:
Combining tunable transcription with an enzyme-degradation tag affords an effective means to reduce intracellular enzyme concentrations from high to very low levels. Such fine-tuned control allows selection pressure to be systematically increased in directed-evolution experiments. This facilitates identification of mutants with wild-type activity, as shown here for an engineered chorismate mutase. Numerous selection formats and cell-based screening methodologies may benefit from the large dynamic range afforded by this easily implemented strategy.
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