来自元基因组图书馆的工程直角翻译系统扩展了遗传密码
Kosuke Seki1, Michael T A Nguyen2,3, Petar I Penev4,5
1Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, USA.
bioRxiv : the preprint server for biology
|November 24, 2025
概括
科学家们从元基因组数据中发现了一种新的正交翻译系统,将非正规氨基酸 (ncAAs) 纳入蛋白质中. 这个系统使遗传密码扩展成为可能,进步了蛋白质的工程和设计.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 合成生物学 合成生物学
背景情况:
- 使用非正规氨基酸 (ncAAs) 的遗传密码扩展增强了蛋白质的功能和设计可能性.
- 目前用于发现正交氨基-tRNA合成酶 (aaRS) 和tRNA对的方法受到低吞吐量和少量可用系统的限制.
研究的目的:
- 发现,描述和设计用于基因代码扩展的新型直角翻译系统.
- 开发一个集成的计算和实验管道,以有效地发现aARS:tRNA对.
- 为了使特定的ncAAs,如5-基,能够被纳入UGA编码子中的蛋白质.
主要方法:
- 超基因组数据挖掘,以确定潜在的正交tRNA和aaRS候选者.
- 通过对超过1,250个aaRS:tRNA组合进行高通量选.
- 在无细胞和细胞环境中鉴别的直角系统的表征.
- 在基因组重新编码的细菌菌株 (Ochre E. coli) 中对特定的ncAA结合的aARS变体的工程.
主要成果:
- 发现和描述AP1TrpRS:tRNATrpUCA正交对,该对与托解码UGA编码子.
- 在无细胞和细胞环境中展示AP1系统的高活性.
- 成功设计了一种AP1 TrpRS变体,用于在Ochre E. coli中的UGA停止码子中加入5-基基.
结论:
- 基因组生物探测,无细胞查和基于细胞的工程的综合管道显著加快了正交转化系统的发现和优化.
- 这项工作扩大了遗传密码扩展工具包,为新型蛋白质设计和功能铺平了道路.
- 开发的战略为合成生物学和蛋白质工程的未来进步提供了一个强大的平台.
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