RS/tRNA Foundational Publication Support
Mehl, Ryan A, Christopher Anderson, Stephen W Santoro, Lei Wang, Andrew B Martin, David S King, David M Horn, and Peter G Schultz. (2003) 2003. “Generation Of A Bacterium With A 21 Amino Acid Genetic Code.”. Journal Of The American Chemical Society 125 (4): 935-9.
RS/tRNA Pair Development Year
2003
ncAA(s) Incorporated
p-Aminophenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
π-donating effects, hydrogen-bonding properties, and weak basicity
RS Organism of Origin
Parent RS
RS Mutations
Y32T
E107T
D158P
I159L
L162A
E107T
D158P
I159L
L162A
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
Other tRNA Mutations
G37A
T17aA
A38C
T45T
T47G
T17aA
A38C
T45T
T47G
RS/tRNA Availability
Can be obtained from AddGene Plasmid #85502
Used in what cell line?
RS/tRNA Additional Notes
In DH10B,
TAG stop codon was introduced at the fourth codon of the sperm whale myoglobin gene to incorporate the nonstandard amino acid para-azidophenylalanine (pAF). In the presence of 1 mM pAF or a biosynthetic pathway for pAF production, full-length myoglobin was expressed, whereas no myoglobin was detected by SDS-PAGE without pAF supplementation. The yield of modified myoglobin (3.0 mg/L) was similar to that of the wild-type (3.5 mg/L), indicating efficient suppression of the TAG codon by the pAF-specific translation system. N-terminal sequencing confirmed that only a nonstandard amino acid, and none of the 20 natural amino acids, occupied the fourth position. Mass spectrometry further validated the incorporation of pAF, with an average protein mass of approximately 18,430 Da, consistent across both exogenous and biosynthetic pAF sources.
TAG stop codon was introduced at the fourth codon of the sperm whale myoglobin gene to incorporate the nonstandard amino acid para-azidophenylalanine (pAF). In the presence of 1 mM pAF or a biosynthetic pathway for pAF production, full-length myoglobin was expressed, whereas no myoglobin was detected by SDS-PAGE without pAF supplementation. The yield of modified myoglobin (3.0 mg/L) was similar to that of the wild-type (3.5 mg/L), indicating efficient suppression of the TAG codon by the pAF-specific translation system. N-terminal sequencing confirmed that only a nonstandard amino acid, and none of the 20 natural amino acids, occupied the fourth position. Mass spectrometry further validated the incorporation of pAF, with an average protein mass of approximately 18,430 Da, consistent across both exogenous and biosynthetic pAF sources.