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A codon optimization method and application for in vitro expression of heterologous genes

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: 2021-08-17
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Claims
  • Application Information

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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

Method used

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  • A codon optimization method and application for in vitro expression of heterologous genes
  • A codon optimization method and application for in vitro expression of heterologous genes
  • A codon optimization method and application for in vitro expression of heterologous genes

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Embodiment 1

[0038] A codon optimization method for the expression of heterologous genes, the specific steps are as follows:

[0039] (1) Get the nucleotide sequence of the host cell allocarin, the correctness of the nucleotide sequence is detected, and the incorrect nucleotide sequence is removed to obtain a nucleotide sequence library for step (2);

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

[0041] (a) whether or not the length of the nucleotide sequence of the various genes complies with 3 multiplexes;

[0042] (b) whether each codon in the gene encoding sequence corresponding to the amino acid sequences of each protein meets the standard codon table;

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

[0044] (2) The statistical formula is as follows: The statistical formula ...

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Abstract

The invention discloses a codon optimization method for in vitro expression of heterologous genes. The method comprises: obtaining the nucleotide sequence of the whole genome of the host cell and the amino acid sequence of the whole protein group; taking the codon pair as the statistical object, and counting The weight of each codon pair in the whole genome of the host cell; select the protein to be optimized, and construct a one-way graph model with codons as nodes and the weight values ​​between upstream and downstream codon pairs as line values; according to the one-way graph Model to obtain the nucleotide sequence of the optimized gene. The present invention uses the whole genome and whole protein group of the host cell as the sequence library, takes codon pairs as statistical objects, and constructs a one-way graph model with codons as nodes and the weight values ​​between upstream and downstream codon pairs as line values, To obtain the optimal codon combination sequence, an optimized gene with an optimized nucleotide sequence can be obtained, and the optimized gene can be highly expressed in vitro, and the expression level is significantly increased.

Description

Technical field [0001] The present invention relates to the field of molecular biology genetic engineering, and more particularly to a codon optimization method for use in heterologous genes, and an optimized hydrothermal green fluorescent protein by this method in a tobacco-free recombinant protein expression system. Background technique [0002] The codon refers to a trim to determine the adjacent three bases of an amino acid in the RNA chain, also known as tri-strain codons. Since the genetic codon has a degenticity, a amino acid is usually compiled by 2 to 6 codons, but the use of the codon is not uniform, nor random, in each species, for the use of the synonym Have preference. [0003] Cryptographic preference analysis is also important in the application of transgenic experiments in the application of transgenic experiments in addition to evolution of species. External genes should be highly expressed in the host and must be optimized for host species codons. Many aspects i...

Claims

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Application Information

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Patent Type & Authority Patents(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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