英国剑桥大学Jason W. Chin研究组实现对正交氨酰-tRNA合成酶与tRNA对的快速发现和进化。这一研究成果于2020年4月13日在线发表在国际学术期刊《自然—生物技术》上。
据研究人员介绍,通过扩增细胞的遗传密码来把非经典的氨基酸整合到蛋白质中存在一个关键的难点,即发现更多的氨酰-tRNA合成酶(aaRS)与tRNA对(在氨酰化特异性上是正交的)。
Title: Rapid discovery and evolution of orthogonal aminoacyl-tRNA synthetase–tRNA pairs
Author: Daniele Cervettini, Shan Tang, Stephen D. Fried, Julian C. W. Willis, Louise F. H. Funke, Lucy J. Colwell, Jason W. Chin
Issue&Volume: 2020-04-13
Abstract: A central challenge in expanding the genetic code of cells to incorporate noncanonical amino acids into proteins is the scalable discovery of aminoacyl-tRNA synthetase (aaRS)–tRNA pairs that are orthogonal in their aminoacylation specificity. Here we computationally identify candidate orthogonal tRNAs from millions of sequences and develop a rapid, scalable approach—named tRNA Extension (tREX)—to determine the in vivo aminoacylation status of tRNAs. Using tREX, we test 243 candidate tRNAs in Escherichia coli and identify 71 orthogonal tRNAs, covering 16 isoacceptor classes, and 23 functional orthogonal tRNA–cognate aaRS pairs. We discover five orthogonal pairs, including three highly active amber suppressors, and evolve new amino acid substrate specificities for two pairs. Finally, we use tREX to characterize a matrix of 64 orthogonal synthetase–orthogonal tRNA specificities. This work expands the number of orthogonal pairs available for genetic code expansion and provides a pipeline for the discovery of additional orthogonal pairs and a foundation for encoding the cellular synthesis of noncanonical biopolymers.
DOI: 10.1038/s41587-020-0479-2
Source: https://www.nature.com/articles/s41587-020-0479-2
Nature Biotechnology:《自然—生物技术》,创刊于1996年。隶属于施普林格·自然出版集团,最新IF:68.164
官方网址:https://www.nature.com/nbt/
投稿链接:https://mts-nbt.nature.com/cgi-bin/main.plex
本期文章:《自然—生物技术》:Online/在线发表