Total synthesis of Escherichia coli with a recoded genome
Total synthesis of Escherichia coli with a recoded genome
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DOI：
10.1038/s41586-019-1192-5
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发表时间：
2019-05-23
期刊：
影响因子：
64.8
通讯作者：
Chin, Jason W.
中科院分区：
文献类型：
--
作者：
Fredens, Julius;Wang, Kaihang;Chin, Jason W.
大自然使用密码子来编码基因组中蛋白质的合成，并从多达6个同义词中选择1个正义密码子来编码每种氨基酸。同义密码子选择有多种重要作用，许多同义替换是有害的。在这里，我们证明了用于编码典型氨基酸的密码子的数量可以通过用定义的同义词在全基因组范围内替换目标密码子来减少。我们通过高保真收敛的全合成创造了一种具有4兆碱基合成基因组的大肠杆菌变种。我们的合成基因组实施了一个明确的重新编码和重构计划--只在七个位置进行简单的更正--以取代基因组中每一个已知的两个正义密码子和一个终止密码子的出现。因此，我们重新编码了18,214个密码子，创建了一个具有61个密码子基因组的有机体；该有机体使用59个密码子来编码20个氨基酸，并能够删除以前必不可少的转移RNA。
Nature uses 64 codons to encode the synthesis of proteins from the genome, and chooses 1 sense codon-out of up to 6 synonyms-to encode each amino acid. Synonymous codon choice has diverse and important roles, and many synonymous substitutions are detrimental. Here we demonstrate that the number of codons used to encode the canonical amino acids can be reduced, through the genome-wide substitution of target codons by defined synonyms. We create a variant of Escherichia coli with a four-megabase synthetic genome through a high-fidelity convergent total synthesis. Our synthetic genome implements a defined recoding and refactoring scheme-with simple corrections at just seven positions-to replace every known occurrence of two sense codons and a stop codon in the genome. Thus, we recode 18,214 codons to create an organism with a 61-codon genome; this organism uses 59 codons to encode the 20 amino acids, and enables the deletion of a previously essential transfer RNA.