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 Protocols and Structural Information
Zhu, Phillip, Ryan A Mehl, and Richard B Cooley. (2023) 2023. “Biosynthesis And Genetic Encoding Of Non-Hydrolyzable Phosphoserine Into Recombinant Proteins In Escherichia Coli.”. Bio-Protocol 13 (21): e4861. doi:10.21769/BioProtoc.4861.
RS/tRNA Pair Development Year
2023
ncAA(s) Incorporated
2-amino-4-phosphonobutanoic acid (nhpSer)
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Used to produce proteins that mimic site-specific phosphorylation at a serine residue - where the phosphate group cannot by hydrolyzed - via genetic code expansion versus other protein phosphorylation strategies.
RS Organism of Origin
Parent RS
RS Mutations
E412P
E414F
P495M
I496W
F529S
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
Entire kit for the "PermaPhos" system (nhpSer incorporation using GCE) available as Addgene kit #1000000226.
Component Addgene IDs are:
201922 - machinery plasmid, including RS/tRNA pair
174075 and 174076 - plasmids for control sfGFP forms which can also be used for other POI
201923 - plasmid for Frb pathway
197656 - E. coli strain BL21(DE3) ΔserC
Component Addgene IDs are:
201922 - machinery plasmid, including RS/tRNA pair
174075 and 174076 - plasmids for control sfGFP forms which can also be used for other POI
201923 - plasmid for Frb pathway
197656 - E. coli strain BL21(DE3) ΔserC
Used in what cell line?
RS/tRNA Additional Notes
While this pair was used for nhpSer incorporation in the Zhu et al. 2023 paper (and therefore listed as "Development Year"), the SepRS2 was developed in 2015, and the Sep-tRNA-v2 was developed in 2017.