An unnatural base pair for incorporating amino acid analogs into proteins

被引：195
作者：
Hirao, I
Ohtsuki, T
Fujiwara, T
Mitsui, T
Yokogawa, T
Okuni, T
Nakayama, H
Takio, K
Yabuki, T
Kigawa, T
Kodama, K
Yokogawa, T
Nishikawa, K
Yokoyama, S
机构：
[1] RIKEN, JST, ERATO, Yokoyama CytoLog Project, Wako, Saitama 3510198, Japan
[2] RIKEN, Genom Sci Ctr, Tsurumi Ku, Yokohama, Kanagawa 2300045, Japan
[3] RIKEN, Biomol Characterizat Div, Wako, Saitama 3510198, Japan
[4] Univ Tokyo, Grad Sch Sci, Dept Biophys & Biochem, Bunkyo Ku, Tokyo 1130033, Japan
[5] Gifu Univ, Fac Engn, Dept Biomol Sci, Gifu 5011193, Japan
来源：
NATURE BIOTECHNOLOGY | 2002年 / 20卷 / 02期
关键词：
D O I：
10.1038/nbt0202-177
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
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.
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页码：177 / 182
页数：6
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