RS/tRNA Foundational Publication Support
Wang, L, A Brock, B Herberich, and P Schultz. (2001) 2001. “Expanding The Genetic Code Of Escherichia Coli.”. Science (New York, N.y.) 292 (5516): 498-500.
RS/tRNA Usage Publications
Chatterjee, Abhishek, Han Xiao, Michael Bollong, Hui-Wang Ai, and Peter G Schultz. (2013) 2013. “Efficient Viral Delivery System For Unnatural Amino Acid Mutagenesis In Mammalian Cells.”. Proceedings Of The National Academy Of Sciences Of The United States Of America 110 (29): 11803-8. doi:10.1073/pnas.1309584110.
Anderson, Christopher, Ning Wu, Stephen W Santoro, Vishva Lakshman, David S King, and Peter G Schultz. (2004) 2004. “An Expanded Genetic Code With A Functional Quadruplet Codon.”. Proceedings Of The National Academy Of Sciences Of The United States Of America 101 (20): 7566-71.
RS/tRNA Pair Development Year
2013
ncAA(s) Incorporated
m-Acetyl-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Labelling with
hydrazides, both in
vitro and on cell
surfaces
hydrazides, both in
vitro and on cell
surfaces
p-azido-L-phenylalanine (pAzF)
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Used as a photocrosslinker, allowing for crosslinking and bioorthogonal ligation of protein.
p-Iodo-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Mutant GFPexcitation/ emission, later used for protein crystallization and as reactive handle. Structural determination of proteins. Amber suppression in a TB model mycobacterium
p-Bromo-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Mutant GFP
excitation/ emission
excitation/ emission
2-Nitro-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
n/a
2-Cyano-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Potentially useful as fluorescence-based environmental sensor to study protein conformation. Also, can be an IR probe.
O-propargyl-L-tyrosine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Reactive handle for azide-alkyne cycloaddition
RS Organism of Origin
Parent RS
RS Mutations
Tyr32Gln
Asp158Ala
Glu107Thr
Leu162Pro
Asp158Ala
Glu107Thr
Leu162Pro
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
RS/tRNA Availability
Can be purchased from Addgene Plasmid #50831. Contains eGFP in the plasmid as well.
RS/tRNA Additional Notes
OMeYRS has been with 1mM of tBoc-lysine in HEK293 cells, which showed expression only in the presence of the ncAA. ~80% of cells were found to express eGFP in the presence of ncAA, while <5% were flourescent in the absence. When tested with 1mM O-Methyl-L-tyrosine in HEK293 cells, a purified yield of ~36ug per 10^6 cells of eGFP with C-terminal His-Tag was observed after isolation with Ni-NTA. It was also shown capable of incorporating 8 other ncAA in mammalian cells when in the pressence of an excess of reporter gene expression.
It has also been tested in cell lines where it has shown high levels of eGFP in the presence of ncAA ( >50% for most, ~5% for mouse embryonic stem cells)
** The plasmid present in Addgene has used two different tRNAcuaTyr from Bacillus stearothermophilus and E.coli.
It has also been tested in cell lines where it has shown high levels of eGFP in the presence of ncAA ( >50% for most, ~5% for mouse embryonic stem cells)
** The plasmid present in Addgene has used two different tRNAcuaTyr from Bacillus stearothermophilus and E.coli.