Artificial translation system using dual sense codon and divided codon boxes

我们设计了“双重遗传密码”和“人工分割密码子盒”两种新的遗传密码重编程策略，使我们能够创建高度工程化的翻译系统。双重遗传密码是由两个不同的重编程遗传密码控制的翻译-解码系统。一种是通常用于延伸反应的遗传密码，另一种是专门用于起始步骤的遗传密码。也就是说，单个密码子具有“双重意义”，在起始和延伸步骤中指定两种不同的氨基酸。在人工分割密码子盒的方法中，一个天然氨基酸占据的密码子盒实际上被分裂，两个不同的氨基酸被重新分配到分裂后的密码子盒的每个部分。这使我们能够指定一个密码子盒用于原始的蛋白质生成氨基酸和另一个所需的非蛋白质生成氨基酸。我们通过实验证明了这两种方法。此外，这两种新策略的整合也进行了，使我们能够彻底重新编程翻译系统的解码规则，涉及广泛的非标准和蛋白质生成构建块（总共超过28个物种）。

We have designed two novel strategies in genetic code reprogramming,"dual genetic code" and "artificial division of codon boxes", enabling us to create extremely engineered translation system. Dual genetic code is translation-decoding system governed by two distinct reprogrammed genetic codes. One is the genetic code used for elongation reaction as usual, and the other is one exclusively used in initiation step. That is, a single codon has" dual sense "designating two distinct amino acids in initiation and elongation steps. In the approach of artificial division of codon boxes, a codon box occupied by one amino acid in natural is virtually split and two distinct amino acids are reassigned into each part of the divided codon box. This enables us to designate a codon box for an original proteinogenic and another desirable non-proteinogenic amino acids. We have experimentally demonstrated these two methodologies. Moreover, integration of these two novel strategies was also carried out, allowing us to drastically reprogram the decoding rule of translation system involving a wide array of non-standard and proteinogenic building blocks(totally over 28 spicies)