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- Title
An unnatural base pair for incorporating amino acid analogs into proteins.
- Authors
Hirao, Ichiro; Ohtsuki, Takashi; Fujiwara, Tsuyoshi; Mitsui, Tsuneo; Yokogawa, Tomoko; Okuni, Taeko; Nakayama, Hiroshi; Takio, Koji; Yabuki, Takashi; Kigawa, Takanori; Kodama, Koichiro; Yokogawa, Takashi; Nishikawa, Kazuya; Yokoyama, Shigeyuki
- Abstract
An unnatural base pair of 2-amino-6-(2-thienyl)purine (denoted by s) and pyridin-2-one (denoted by y) was developed to expand the genetic code. The ribonucleoside triphosphate of y was site-specifically incorporated into RNA, opposite s in a template, by T7 RNA polymerase. This transcription was coupled with translation in an Escherichia coli cell-free system. The yAG codon in the transcribed ras mRNA was recognized by the CUs anticodon of a yeast tyrosine transfer RNA (tRNA) variant, which had been enzymatically aminoacylated with an unnatural amino acid, 3-chlorotyrosine. Site-specific incorporation of 3-chlorotyrosine into the Ras protein was demonstrated by liquid chromatography-mass spectrometry (LC-MS) analysis of the products. This coupled transcription-translation system will permit the efficient synthesis of proteins with a tyrosine analog at the desired position.
- Publication
Nature biotechnology, 2002, Vol 20, Issue 2, p177
- ISSN
1087-0156
- Publication type
Journal Article
- DOI
10.1038/nbt0202-177