PDF(Pubmed)
Abstract：
Manipulating transfer RNAs (tRNAs) for emancipating sense codons to simplify genetic codons in a cell-free protein synthesis (CFPS) system can offer more flexibility and controllability. Here, we provide an overview of the tRNA complement protein synthesis system construction in the tRNA-depleted Protein synthesis Using purified Recombinant Elements (PURE) system or S30 extract. These designed polypeptide coding sequences reduce the genetic codon and contain only a single tRNA corresponding to a single amino acid in this presented system. Strategies for removing tRNAs from cell lysates and synthesizing tRNAs in vivo/vitro are summarized and discussed in detail. Furthermore, we point out the trend toward a minimized genetic codon for reducing codon redundancy by manipulating tRNAs in the different proteins. It is hoped that the tRNA complement protein synthesis system can facilitate the construction of minimal cells and expand the biomedical application scope of synthetic biology.
摘要：
在无细胞蛋白质合成（CFPS）系统中，操纵转移RNA（tRNA）以解放有义密码子以简化遗传密码子可以提供更多的灵活性和可控性。这里,我们提供了使用纯化的重组元件（PURE）系统或S30提取物在tRNA耗尽的蛋白质合成中构建tRNA补体蛋白合成系统的概述。这些设计的多肽编码序列减少了遗传密码子,并且仅含有对应于该系统中单个氨基酸的单个tRNA。总结并详细讨论了从细胞裂解物中去除tRNA和体内/体外合成tRNA的策略。此外，我们指出了通过操纵不同蛋白质中的tRNA来减少密码子冗余的最小化遗传密码子的趋势。希望tRNA补体蛋白合成系统能够促进最小细胞的构建，扩大合成生物学的生物医学应用范围。
