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A Facile System for Genetic Incorporation of Two Different Noncanonical Amino Acids into One Protein in Escherichia coli


  • We thank Prof. Peter G. Schultz of The Scripps Research Institute for providing us with the pEVOL-AzFRS plasmid and Prof. Daniel Romo, Prof. Kevin Burgess, and Prof. Jiong Yang of Texas A&M University for using their instruments. This work was supported by Welch Grant A-1715 and the New Faculty Startup Fund to W.R.L. from the Chemistry Department of Texas A&M University.


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Two's company: Using a wild-type or evolved PylRS-pylTUUA pair to suppress ochre mutation and an evolved MjTyrRS-Mjequation image pair to suppress amber mutation, two different noncanonical amino acids (NAAs) have been concomitantly incorporated into one protein in E. coli with high efficiency (see picture, with NAAs 14; GFP=green-fluorescent protein).

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