RIKEN Systems and Structural Biology Center, Tsurumi, Yokohama, Japan.
Biochem Biophys Res Commun. 2011 Aug 12;411(4):757-61. doi: 10.1016/j.bbrc.2011.07.020. Epub 2011 Jul 18.
The genetic encoding of synthetic or "non-natural" amino acids promises to diversify the functions and structures of proteins. We applied rapid codon-reassignment for creating Escherichia coli strains unable to terminate translation at the UAG "stop" triplet, but efficiently decoding it as various tyrosine and lysine derivatives. This complete change in the UAG meaning enabled protein synthesis with these non-natural molecules at multiple defined sites, in addition to the 20 canonical amino acids. UAG was also redefined in the E. coli BL21 strain, suitable for the large-scale production of recombinant proteins, and its cell extract served the cell-free synthesis of an epigenetic protein, histone H4, fully acetylated at four specific lysine sites.
遗传编码合成或“非天然”氨基酸有望使蛋白质的功能和结构多样化。我们应用快速密码子重排技术，创建了不能在 UAG“终止”三联体处终止翻译但能有效解码为各种酪氨酸和赖氨酸衍生物的大肠杆菌菌株。这种 UAG 含义的完全改变，使得除了 20 种标准氨基酸外，还可以在多个定义的位点上用这些非天然分子进行蛋白质合成。UAG 还在适合重组蛋白大规模生产的大肠杆菌 BL21 菌株中重新定义，其细胞提取物用于在无细胞体系中合成组蛋白 H4 的完全乙酰化的四个特定赖氨酸位点。
