A Rationally Designed Aminoacyl-tRNA Synthetase for Genetically Encoded Fluorescent Amino Acids
2016
The incorporation of non-canonical amino acids into proteins has emerged as a promising strategy to
manipulate and study protein structure-function relationships with superior precision in vitro and in vivo. To date,
fluorescent non-canonical amino acids (f-ncAA) have been successfully incorporated in proteins expressed in bacterial
systems, Xenopus oocytes, and HEK-293T cells. Here, we describe the rational generation of an orthogonal aminoacyltRNA
synthetase based on the E. coli tyrosine synthetase that is capable of encoding the f-ncAA tyr-coumarin in HEK-293T
cells.
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