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Updated: Jul 9, 2025

OnePot PURE Cell-Free System
Published on: June 23, 2021
Changes in Coding and Efficiency through Modular Modifications to a One Pot PURE System for In Vitro Transcription
Phuoc H T Ngo1, Satoshi Ishida1, Bianca B Busogi1
1Department of Molecular Biosciences, University of Texas at Austin, Austin, Texas 78712, United States.
Abstract:
The incorporation of unnatural amino acids is an attractive method for improving or bringing new and novel functions in peptides and proteins. Cell-free protein synthesis using the Protein Synthesis Using Recombinant Elements (PURE) system is an attractive platform for efficient unnatural amino acid incorporation. In this work, we further adapted and modified the One Pot PURE to obtain a robust and modular system for enzymatic single-site-specific incorporation of an unnatural amino acid. We demonstrated the flexibility of this system through the introduction of two different orthogonal aminoacyl tRNA synthetase:tRNA pairs that suppressed two distinctive stop codons in separate reaction mixtures.
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