Authors
Yuki Goto, Takayuki Katoh, and Hiroaki Suga
Summary
Genetic code reprogramming is a method for the reassignment of arbitrary codons from proteinogenic amino acids to non-proteinogenic ones, and thus specific sequences of non-standard peptides can be ribosomally expressed according to their mRNA templates. We have developed a protocol that facilitates the genetic code reprogramming using flexizymes integrated with a custom-made in-vitro translation apparatus, referred to as the flexible in-vitro translation (FIT) system. Flexizymes are flexible tRNA acylation ribozymes that enable the preparation of a diverse array of non-proteinogenic acyl-tRNAs. These acyl-tRNAs read vacant codons created in the FIT system, yielding the desired non-standard peptides with diverse exotic structures, such as N-methyl amino acids, D-amino acids and physiologically stable macrocyclic scaffolds. We here describe a protocol for the preparation of DNA templates for flexizymes, tRNAs, microhelix RNA, and mRNAs that are used for genetic code reprogramming with flexizymes.Further details
The protocol was published on Protocol Exchange on 12 May 2011. To see the entire protocol, click on the source link.Advertisement
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