RS/tRNA Pair Development Year
2019
RS Organism of Origin
Parent RS
RS Mutations
P186T
L274R
N311G
C313N
L274R
N311G
C313N
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
RS/tRNA Availability
The codon optimized RS and tRNA are available on a pAcBac plasmid as Addgene #141173
RS/tRNA Additional Notes
In E coli, this RS was said to perform about as well as the "5B" nitroTyr MjRS. It generated full-fidelity 3NY incorporation into sfGFP(150) at a yield of ~140 mg/L (~30% efficiency) and had a very low UP50 of ~3 microM. Mn-superoxide dismutase(34) and 14-3-3(130) were also produced in E coli. For use in mammalian cells, adding a nuclear export signal (NES) to the RS improve its efficiency by a factor of ~1.8. As 3NY was toxic to HEK cells at 1 mM, 0.3 mM was used for these expressions. MS analysis showed full-fidelity incorporation into sfGFP(150), and MnSOD(34) and 14-3-3(130) were expressed at ~10-30% efficiencies.