Using a solid-phase ribozyme aminoacylation system to reprogram the genetic code.

Hiroshi Murakami; Dimitrios Kourouklis; Hiroaki Suga

doi:10.1016/j.chembiol.2003.10.010

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Using a solid-phase ribozyme aminoacylation system to reprogram the genetic code.

Hiroshi Murakami·Dimitrios Kourouklis·Hiroaki Suga

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Abstract

Here, we report a simple and economical tRNA aminoacylation system based upon a resin-immobilized ribozyme, referred to as Flexiresin. This catalytic system features a broad spectrum of activities toward various phenylalanine (Phe) analogs and suppressor tRNAs. Most importantly, it allows users to perform the tRNA aminoacylation reaction and isolate the aminoacylated tRNAs in a few hours. We coupled the Flexiresin system with a high-performance cell-free translation system and demonstrated protein mutagenesis with seven different Phe analogs in parallel. Thus, the technology developed herein provides a new tool that significantly simplifies the procedures for the synthesis of aminoacyl-tRNAs charged with nonnatural amino acids, which makes the nonnatural amino acid mutagenesis of proteins more user accessible.

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Acylation Base Sequence Catalysis Cell-Free System DNA Primers Genetic Code Mutagenesis, Site-Directed Protein Processing, Post-Translational RNA, Catalytic RNA, Transfer, Asn Journal Article Research Support, Non-U.S. Gov't

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Hiroshi Murakami, Dimitrios Kourouklis, Hiroaki Suga. 2003. Using a solid-phase ribozyme aminoacylation system to reprogram the genetic code.. https://doi.org/10.1016/j.chembiol.2003.10.010

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