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

A technology of codon optimization and in vitro expression, applied in peptide sources, instruments, peptides, etc., can solve the problem of low expression of exogenous genes and achieve the effect of increasing expression

Active Publication Date: 2019-11-22
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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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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  • Codon optimization method used for heterologous gene in-vitro expression and application
  • Codon optimization method used for heterologous gene in-vitro expression and application
  • Codon optimization method used for heterologous gene in-vitro expression and application

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Embodiment 1

[0038] A codon optimization method for heterologous gene expression in vitro, the specific steps are as follows:

[0039] (1) Obtain the nucleotide sequence of the whole genome of the host cell, detect the correctness of the nucleotide sequence, remove the incorrect nucleotide sequence, and obtain the nucleotide sequence library used for step (2) statistics;

[0040] The method for detecting the correctness of the nucleotide sequence is:

[0041] (a) compare whether the length of the nucleotide sequence of each gene conforms to a multiple of 3;

[0042] (b) compare whether each codon in the gene coding sequence corresponding to the amino acid sequence of each protein conforms to the standard codon table;

[0043] If (a) and (b) are both consistent, it is determined that the nucleotide sequence and amino acid sequence are correct; in other cases, it is determined that the nucleotide sequence and amino acid sequence are incorrect.

[0044] (2) Taking the codon pair as the stat...

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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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G16B25/10G16B30/10C07K14/435
CPCC07K14/43595G16B25/10G16B30/10
Inventor 徐盛春郑元庭徐飞李素娟邵健丰王钢军李琪恺
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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