2000
DOI: 10.1021/ja000595y
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A New Functional Suppressor tRNA/Aminoacyl−tRNA Synthetase Pair for the in Vivo Incorporation of Unnatural Amino Acids into Proteins

Abstract: General methods for selectively incorporating unnatural amino acids into proteins in vivo, directly from the growth media, would greatly expand our ability to manipulate protein structure and function. 1 For example, the ability to place fluorophores selectively into proteins in vivo would provide powerful tools for cell biology, or the ability to generate large quantities of proteins with metal binding or keto amino acids might lead to proteins with enhanced physical or catalytic properties. Our approach invo… Show more

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Cited by 143 publications
(145 citation statements)
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“…Besides providing the only example of a 21st synthetasetRNA pair for use in a eukaryotic system, our approach differs somewhat from that of Schultz and coworkers, in which both the aaRS and the suppressor tRNA are imported from a heterologous organism (13,14), and it offers certain advantages. For example, because the suppressor tRNAs used are derived from tRNA of the same organism or a related organism, transcripts derived from the suppressor tRNA genes are likely to be processed well to yield functional suppressor tRNAs (34).…”
Section: Discussionmentioning
confidence: 99%
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“…Besides providing the only example of a 21st synthetasetRNA pair for use in a eukaryotic system, our approach differs somewhat from that of Schultz and coworkers, in which both the aaRS and the suppressor tRNA are imported from a heterologous organism (13,14), and it offers certain advantages. For example, because the suppressor tRNAs used are derived from tRNA of the same organism or a related organism, transcripts derived from the suppressor tRNA genes are likely to be processed well to yield functional suppressor tRNAs (34).…”
Section: Discussionmentioning
confidence: 99%
“…The approach used by Schultz and coworkers (13,14), in which both the aaRS and the suppressor tRNA are imported from a heterologous organism, is based on the inability of a few eubacterial aaRSs to aminoacylate the corresponding tRNAs from eukaryotes or archaebacteria and vice versa. For example, the E. coli GlnRS does not aminoacylate yeast tRNA Gln , and the yeast GlnRS does not aminoacylate E. coli tRNA Gln (37).…”
Section: Discussionmentioning
confidence: 99%
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“…Notably, the M. jannaschii TyrRS does not recognize any E. coli tRNA, and the suppressor tRNA is not recognized by any E. coli aaRS. The employment of such a tRNA⅐aaRS pair that is ''orthogonal'' to the host system (23)(24)(25), along with the engineering of the aaRS amino acid specificity, allows the reacylation of the tRNA with the unnatural amino acid This paper was submitted directly (Track II) to the PNAS office.…”
mentioning
confidence: 99%
“…Recently, two cognate pairs were developed for use in intact Escherichia coli cells (23,24). One pair is specific for glutamine and is comprised of a yeast tRNA and synthetase.…”
mentioning
confidence: 99%