RS/tRNA Foundational Publication Support
Liu, Chang C, and Peter G Schultz. (2006) 2006. “Recombinant Expression Of Selectively Sulfated Proteins In Escherichia Coli.”. Nature Biotechnology 24 (11): 1436-40.
RS/tRNA Usage Publications
Liu, Chang C, Susan E Cellitti, Bernhard H Geierstanger, and Peter G Schultz. (2009) 2009. “Efficient Expression Of Tyrosine-Sulfated Proteins In E. Coli Using An Expanded Genetic Code.”. Nature Protocols 4 (12): 1784-9. doi:10.1038/nprot.2009.188.
Schwessinger, Benjamin, Xiang Li, Thomas L Ellinghaus, Leanne Jade G Chan, Tong Wei, Anna Joe, Nicholas Thomas, et al. (2016) 2016. “A Second-Generation Expression System For Tyrosine-Sulfated Proteins And Its Application In Crop Protection.”. Integrative Biology : Quantitative Biosciences From Nano To Macro 8 (4): 542-5. doi:10.1039/c5ib00232j.
RS/tRNA Pair Development Year
2006
ncAA(s) Incorporated
Sulfo-L-tyrosine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Labelling of muscles
(global)
Dipeptide
(DanAla-‐Ala)
required for
uptake and
bioavailability
(global)
Dipeptide
(DanAla-‐Ala)
required for
uptake and
bioavailability
RS Organism of Origin
Parent RS
RS Mutations
Tyr32Leu,
Leu65Pro
Asp158Gly
Ile159Cys
Leu162Lys
Leu65Pro
Asp158Gly
Ile159Cys
Leu162Lys
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
RS/tRNA Availability
Can be obtained from AddGene Plasmid #82417, the plasmid system pULTRA with the STyrRS has been shown to have been shown to higher levels of sY incorporation at single or multiple UAG codons.
RS/tRNA Additional Notes
In C321.ΔA ,
The RS was tested To verify unique incorporation of sulfotyrosine by the selected synthetase STyrRS, an amber mutant (residue 7) of a C-terminal His6 tag Z-domain protein. The incorporation was confirmed by MALDI-TOF.
In DH10B,
Was utilized to express Hirudin from Hirudo medicinalis with TAG 63, at 10mM sulfotyrosine getting a yeild of 5mg/L compared to WT yield being 12 mg/L with incorpotation also being confirmed by MALDI-TOF
The RS was tested To verify unique incorporation of sulfotyrosine by the selected synthetase STyrRS, an amber mutant (residue 7) of a C-terminal His6 tag Z-domain protein. The incorporation was confirmed by MALDI-TOF.
In DH10B,
Was utilized to express Hirudin from Hirudo medicinalis with TAG 63, at 10mM sulfotyrosine getting a yeild of 5mg/L compared to WT yield being 12 mg/L with incorpotation also being confirmed by MALDI-TOF