Codon optimization method used for heterologous gene in-vitro expression and application

Abstract

The invention discloses a codon optimization method used for heterologous gene in-vitro expression. The method comprises the steps that a nucleotide sequence of a whole genome of a host cell and an amino acid sequence of a whole proteome of the host cell are acquired; codon pairs are taken as a statistical object, and statistics is conducted on the weight of each codon pair in the whole genome ofthe host cell; to-be-optimized protein is selected, and a single-direction graph model which takes codons as nodes and takes weight values of the upstream and downstream codon pairs as linear values is constructed; according to the single-direction graph model, a nucleotide sequence of an optimized gene is obtained. According to the codon optimization method used for heterologous gene in-vitro expression, the whole genome and whole proteome of the host cell are utilized as sequence libraries, the codon pairs are taken as the statistical object, and by constructing the single-direction graph model which takes the codons as the nodes and takes the weight values of the upstream and downstream codon pairs as linear values, the optimum codon combination sequence is obtained; the optimized genewith the optimum nucleotide sequence is obtained and can be efficiently expressed in vitro, and the expression amount is significantly improved.

Description

technical field [0001] The invention relates to the technical field of molecular biology genetic engineering, in particular to a codon optimization method for in vitro expression of heterologous genes and the application of the method to obtain optimized jellyfish green fluorescent protein in a tobacco cell-free recombinant protein expression system. Background technique [0002] A codon refers to the three adjacent bases that determine an amino acid on the messenger RNA chain, also known as a triplet codon. Due to the degeneracy of genetic codes, an amino acid is usually encoded by 2 to 6 codons, but the use of codons is not uniform or random. In each species, the use of synonymous codons Preferential. [0003] Codon bias analysis is not only widely used in the evolution of species selection, but also important in the application of transgenic experiments. To efficiently express foreign genes in the host, the codons of the host species must be optimized. Codon optimizati...

AI Technical Summary

Problems solved by technology

However, there are still many problems in the expression of foreign genes in heterologous host cells. Because the common codons of heterologous hosts are far from those of foreign genes, the expression level of foreign genes is relatively low

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