posted on 2023-12-05, 12:04authored byPhuoc
H. T. Ngo, Satoshi Ishida, Bianca B. Busogi, Hannah Do, Maximiliano A. Ledesma, Shaunak Kar, Andrew Ellington
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.