Sisido M, Hohsaka T
Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University, Japan.
Appl Microbiol Biotechnol. 2001 Oct;57(3):274-81. doi: 10.1007/s002530100791.
Position-specific incorporation of nonnatural amino acids into proteins (nonnatural mutagenesis) via an in vitro protein synthesizing system was applied to incorporate a variety of amino acids carrying specialty side groups. A list of nonnatural amino acids thus far successfully incorporated through in vitro translation systems is presented. The position of nonnatural amino acid incorporation was directed by four-base codon/anticodon pairs such as CGGG/CCCG and AGGU/ACCU. The four-base codon strategy was more efficient than the amber codon strategy and could incorporate multiple nonnatural amino acids into single proteins. This multiple mutagenesis will find wide applications, especially in building paths of electron transfer on proteins. The extension of translation systems by the introduction of nonnatural amino acids, four-base codon/anticodon pairs, orthogonal tRNAs, and artificial aminoacyl tRNA synthetases, is a promising approach towards the creation of "synthetic microorganisms" with specialty functions.
通过体外蛋白质合成系统将非天然氨基酸位点特异性地掺入蛋白质中（非天然诱变），被用于掺入各种带有特殊侧链基团的氨基酸。列出了迄今为止通过体外翻译系统成功掺入的非天然氨基酸。非天然氨基酸掺入的位置由四碱基密码子/反密码子对（如CGGG/CCCG和AGGU/ACCU）引导。四碱基密码子策略比琥珀密码子策略更有效，并且可以将多种非天然氨基酸掺入单个蛋白质中。这种多重诱变将有广泛的应用，特别是在构建蛋白质上的电子传递途径方面。通过引入非天然氨基酸、四碱基密码子/反密码子对、正交tRNA和人工氨酰tRNA合成酶来扩展翻译系统，是创建具有特殊功能的“合成微生物”的一种有前景的方法。
