An expanded eukaryotic genetic code
An expanded eukaryotic genetic code
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DOI：
10.1126/science.1084772
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发表时间：
2003-08-15
期刊：
影响因子：
56.9
通讯作者：
Schultz, PG
中科院分区：
文献类型：
--
作者：
Chin, JW;Cropp, TA;Schultz, PG
我们描述了一种将非天然氨基酸添加到酿酒酵母遗传密码中的通用和快速方法。五种氨基酸被高效地和高保真地结合到蛋白质中，以响应无意义的密码子标签。这些氨基酸的侧链包含一个酮基，它可以在体外和体内用各种化学探针和试剂进行独特的修饰；一个用于结构研究的重原子氨基酸；以及用于细胞蛋白质相互作用研究的光交联剂。这种方法不仅消除了遗传密码对我们在酵母中操纵蛋白质结构和功能的能力施加的限制，而且为系统地扩展多细胞真核生物的遗传密码提供了一条途径。
We describe a general and rapid route for the addition of unnatural amino acids to the genetic code of Saccharomyces cerevisiae. Five amino acids have been incorporated into proteins efficiently and with high fidelity in response to the nonsense codon TAG. The side chains of these amino acids contain a keto group, which can be uniquely modified in vitro and in vivo with a wide range of chemical probes and reagents; a heavy atom-containing amino acid for structural studies; and photocrosslinkers for cellular studies of protein interactions. This methodology not only removes the constraints imposed by the genetic code on our ability to manipulate protein structure and function in yeast, it provides a gateway to the systematic expansion of the genetic codes of multicellular eukaryotes.