RS/tRNA Foundational Publication Support
Zhang, Shen, and Hui-Wang Ai. (2020) 2020. “A General Strategy To Red-Shift Green Fluorescent Protein-Based Biosensors.”. Nature Chemical Biology 16 (12): 1434-1439. doi:10.1038/s41589-020-0641-7.
RS/tRNA Pair Development Year
2021
ncAA(s) Incorporated
3-amino-L-tyrosine(NHY)2
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Used to probe radical propagation in ribonucleotide reductase
RS Organism of Origin
Parent RS
RS Mutations
H70S
D158T
I159S
D286Y
D158T
I159S
D286Y
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
RS/tRNA Availability
Can be purchased from AddGene Plasmid #153557
Used in what cell line?
RS/tRNA Additional Notes
In DH10B,
sfGFP containing an amber codon at residue 66 expressed with MjaYRS / tRNA. In the presence of the noncanonical amino acid 2mM aY, they observed strong fluorescence and successfully obtained full-length protein at a yield of ~6.7 mg/L. Without aY, protein production was minimal. Mass spectrometry confirmed the site-specific incorporation of aY, with no evidence of tyrosine misincorporation, demonstrating the specificity of MjaYRS.
sfGFP containing an amber codon at residue 66 expressed with MjaYRS / tRNA. In the presence of the noncanonical amino acid 2mM aY, they observed strong fluorescence and successfully obtained full-length protein at a yield of ~6.7 mg/L. Without aY, protein production was minimal. Mass spectrometry confirmed the site-specific incorporation of aY, with no evidence of tyrosine misincorporation, demonstrating the specificity of MjaYRS.