Expanding the genetic code of Escherichia coli.
A unique transfer RNA (tRNA)/aminoacyl-tRNA synthetase pair has been generated that expands the number of genetically encoded amino acids in Escherichia coli. When introduced into E. coli, this pair leads to the in vivo incorporation of the synthetic amino acid O-methyl-l-tyrosine into protein in response to an amber nonsense codon. The fidelity of translation is greater than 99%, as determined by analysis of dihydrofolate reductase containing the unnatural amino acid. This approach should provide a general method for increasing the genetic repertoire of living cells to include a variety of amino acids with novel structural, chemical, and physical properties not found in the common 20 amino acids.
Wang
L
L
Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Brock
A
A
Herberich
B
B
Schultz
P G
PG
D000926:Anticodon; D003062:Codon / Q000235:genetics / Q000378:metabolism; D018388:Codon, Terminator; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D005815:Genetic Code; D013058:Mass Spectrometry; D017017:Methanococcus / Q000201:enzymology / Q000235:genetics; D008781:Methyltyrosines / Q000378:metabolism*; D009154:Mutation; D014176:Protein Biosynthesis; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism; D012343:RNA, Transfer / Q000235:genetics / Q000378:metabolism*; D012365:RNA, Transfer, Tyr / Q000235:genetics / Q000378:metabolism*; D013489:Suppression, Genetic; D046249:Transfer RNA Aminoacylation; D014169:Transformation, Bacterial; D014448:Tyrosine-tRNA Ligase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D000926:Anticodon; D003062:Codon; D018388:Codon, Terminator; D008781:Methyltyrosines; D012329:RNA, Bacterial; D012365:RNA, Transfer, Tyr; C002350:O-methyltyrosine; D012343:RNA, Transfer; D014448:Tyrosine-tRNA Ligase