RS/tRNA Foundational Publication Support
Lang, Kathrin, Lloyd Davis, Stephen Wallace, Mohan Mahesh, Daniel J Cox, Melissa L Blackman, Joseph M Fox, and Jason W Chin. (2012) 2012. “Genetic Encoding Of Bicyclononynes And Trans-Cyclooctenes For Site-Specific Protein Labeling In Vitro And In Live Mammalian Cells Via Rapid Fluorogenic Diels-Alder Reactions.”. Journal Of The American Chemical Society 134 (25): 10317-20. doi:10.1021/ja302832g.
RS/tRNA Pair Development Year
2012
ncAA(s) Incorporated
endo-BCN-L-Lysine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Reactive handle for SPAAC
Boc-LysOH
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Hydroxy acid to introduce ester linkages into protein backbone
RS Organism of Origin
Parent RS
RS Mutations
Y271M
L274G
C313A
L274G
C313A
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
RS/tRNA Availability
n/a
RS/tRNA Additional Notes
In E.coli, produced 6-12 mg protein per liter into an sfGFP150 construct with incorporation verified both by mass spectrometry and reactivity with tetrazine labels. The RS also incorporated a closely related ncAA with a trans-cyclooctene ring.
In HEK-293 cells, ncAAs were incorporated into an mCherry-eGFP construct, and position 128 of the extracellular EGF receptor, and the nuclear protein jun in a jun-mCherry construct. Both extracellular and nuclear proteins could react with a tetrazine label.
In HEK-293 cells, ncAAs were incorporated into an mCherry-eGFP construct, and position 128 of the extracellular EGF receptor, and the nuclear protein jun in a jun-mCherry construct. Both extracellular and nuclear proteins could react with a tetrazine label.