RS/tRNA Foundational Publication Support
Zhang, Michael Shaofei, Simon F Brunner, Nicolas Huguenin-Dezot, Alexandria D Liang, Wolfgang H Schmied, Daniel T Rogerson, and Jason W Chin. (2017) 2017. “Biosynthesis And Genetic Encoding Of Phosphothreonine Through Parallel Selection And Deep Sequencing.”. Nature Methods 14 (7): 729-736. doi:10.1038/nmeth.4302.
RS/tRNA Pair Development Year
2017
ncAA(s) Incorporated
phosphothreonine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Used to produce site-specifically phosphorylated proteins at a threonine residue via genetic code expansion versus other protein phosphorylation strategies.
RS Organism of Origin
Parent RS
RS Mutations
G320A
L321Y
E412P
E414F
P495M
I496W
F529S
L321Y
E412P
E414F
P495M
I496W
F529S
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
Other tRNA Mutations
C2U
G4C
G5C
G29A
A31U
G34C
C35U
G37A
U39A
C40U
C41U
C67G
C69G
G71A
G4C
G5C
G29A
A31U
G34C
C35U
G37A
U39A
C40U
C41U
C67G
C69G
G71A
RS/tRNA Availability
Addgene #173899 is for the plasmid for pThrRS and PduX (which is a Thr kinase).
Addgene #174075 and #174076 are for control sfGFP WT and 150TAG, respectively. To use these plasmids for a different POI, you can clone in your POI in place of the sfGFP construct. The sfGFP/POI plasmids are not the ones originally used with this system, but work well with it.
Competent cells of BL21(DE3) ΔserC ΔycdX can be purchased commercially from Amid Biosciences (Catalog # BLPCY-201).
Addgene #174075 and #174076 are for control sfGFP WT and 150TAG, respectively. To use these plasmids for a different POI, you can clone in your POI in place of the sfGFP construct. The sfGFP/POI plasmids are not the ones originally used with this system, but work well with it.
Competent cells of BL21(DE3) ΔserC ΔycdX can be purchased commercially from Amid Biosciences (Catalog # BLPCY-201).
Used in what cell line?