In-vitro cell-free protein synthesis system for inserting non-natural amino acid, and method and use

Abstract

Provided is an in-vitro cell-free protein synthesis system for inserting a non-natural amino acid. The reaction system comprises: (1) a yeast cell extracting solution; (2) a non-natural amino acid; (3) an exogenous orthogonal aminoacyl tRNA synthetase/orthogonal tRNA pair; and (4) a template containing a target protein gene sequence, wherein at least one codon encoding an amino acid in the target protein gene sequence is mutated into a stop codon. By means of the reaction system, the insertion efficiency of the non-natural amino acid and the expression quantity of the non-natural amino acid protein can be increased.

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