RS/tRNA Foundational Publication Support
Tanrikulu, Caglar, Emmanuelle Schmitt, Yves Mechulam, William A Goddard, and David A Tirrell. (2009) 2009. “Discovery Of Escherichia Coli Methionyl-Trna Synthetase Mutants For Efficient Labeling Of Proteins With Azidonorleucine In Vivo.”. Proceedings Of The National Academy Of Sciences Of The United States Of America 106 (36): 15285-90. doi:10.1073/pnas.0905735106.
RS/tRNA Usage Publications
Mahdavi, Alborz, Janek Szychowski, John T Ngo, Michael J Sweredoski, Robert L J Graham, Sonja Hess, Olaf Schneewind, Sarkis K Mazmanian, and David A Tirrell. (2014) 2014. “Identification Of Secreted Bacterial Proteins By Noncanonical Amino Acid Tagging.”. Proceedings Of The National Academy Of Sciences Of The United States Of America 111 (1): 433-8. doi:10.1073/pnas.1301740111.
RS/tRNA Pair Development Year
2009
ncAA(s) Incorporated
Azidonorleucine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Biorthogonal Ligation, Click Chemistry
Azidohomoalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
click chemistry, BONCAT (Bioorthogonal non-canonical amino-acid tagging)
RS Organism of Origin
Parent RS
RS Mutations
L13N
Y306L
H301L
Y306L
H301L
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
UAC
RS/tRNA Availability
Can be obtained from Addgene Plasmid #51401
Used in what cell line?
RS/tRNA Additional Notes
In DH10B, the RS is shown to be extremely effective at stringent ncAA concentration. At 0.3mM Anl, NLL appeared to be more than 50% of the clones sampled. Even being the dominate mutant at 0.1mM Anl Concentration. When tested with addtion of Met at varying concentration of 0.01mM, 0.01mM and 0.1mM while in the presence of 1mM Anl, final population showedonly limited sensitivity to Met as deteced through flouresence screening . This was still the case even when Met added at 1.0mM concentration while keeping the concentration of Anl constant of 1mM.
In Y.enterocolitica, when expressing NLL-MetRS constitutively under control of its natural promoter showed the incorporation of Anl in protien detected after being secreted under LCR condition while being tagged with an alkyne-functionalized tetramethylrhodamine (TAMRA) dye and detected by in-gel flouresence labelling.
*Use L274GMm MetRS for Anl Incorporation in mammalian cells*
In Y.enterocolitica, when expressing NLL-MetRS constitutively under control of its natural promoter showed the incorporation of Anl in protien detected after being secreted under LCR condition while being tagged with an alkyne-functionalized tetramethylrhodamine (TAMRA) dye and detected by in-gel flouresence labelling.
*Use L274GMm MetRS for Anl Incorporation in mammalian cells*