Method for designing human aldehyde dehydrogenase 2 sequence according to preferred codon of pichia pastoris

An acetaldehyde dehydrogenase and codon-preferred technology, which is applied in the field of designing gene sequences and can solve the problems of narrow access to acetaldehyde dehydrogenase and the like

Inactive Publication Date: 2011-06-22
WUHAN INSTITUTE OF TECHNOLOGY
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  • Abstract
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Problems solved by technology

In recent years, there have been some reports of anti-alcoholic drugs containing aldehyde dehydrogenase, but the narrow access to aldehyde dehydrogenase has become the bottleneck of this type of health care products

Method used

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  • Method for designing human aldehyde dehydrogenase 2 sequence according to preferred codon of pichia pastoris
  • Method for designing human aldehyde dehydrogenase 2 sequence according to preferred codon of pichia pastoris
  • Method for designing human aldehyde dehydrogenase 2 sequence according to preferred codon of pichia pastoris

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0045] 1. Acquisition and verification of plasmid pPIC9K-ALDH2

[0046] 1. Target sequence design and synthesis:

[0047] The method for designing the sequence of human acetaldehyde dehydrogenase 2 according to the preferred codon of Pichia pastoris, the DNA molecule designed according to the preference of Pichia pastoris, the molecule encodes a protein encoded by the human acetaldehyde dehydrogenase 2 gene gene ID 217 Nucleotide sequences identical at the amino acid level.

[0048] The described human acetaldehyde dehydrogenase 2 gene sequence designed according to the preferred codons of Pichia pastoris is based on the human ALDH2 gene (gene ID 217) [from the human ALDH2 DNA (gene ID217) sequence on GenBank], deleting UTR and Signal peptide and endonuclease EcoR I and endonuclease Not I restriction sites, and then replace the codon GCG encoding Ala with GCU according to the codon usage of Pichia pastoris, and replace the codon CGC and CGG encoding Arg AGA, AGG, CGU and CGA...

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Abstract

The invention relates to a method for designing gene sequences, in particular to a human aldehyde dehydrogenase 2 sequence designed according to a preferred codon of pichia pastoris. The sequence is characterized in that a DNA molecule is designed according to the preference of the pichia pastoris, and a code of the molecule has the same nucleotide sequence as a protein coded by a human aldehyde dehydrogenase 2 gene ID 217 on an amino acid level. The human aldehyde dehydrogenase 2 gene sequence designed according to the preferred codon of pichia pastoris is realized by the following steps: on the basis of a human ALDH2 gene (gene ID 217), UTR and signal peptide are deleted; enzyme cutting sites of an incision enzyme EcoR I and an incision enzyme Not I are added; a codon GCG for coding Ala is replaced by GCU according to the use method of the pichia pastoris codon, and codons CGC and CGG for coding Arg are replaced by AGA, AGG, CGU and CGA; and altogether 28 bases are modified to obtain a target sequence ALDH2. The gene sequence can express with high efficiency in the pichia pastoris, can be applied to preventing alcoholism and protecting livers and has the effects of preventing alcoholism and protecting livers.

Description

technical field [0001] The invention relates to a method for designing gene sequences. Background technique [0002] With the rapid development of the economy and the continuous improvement of people's living standards, people pay more and more attention to high-quality life, the most important of which is health. However, economic development is accompanied by changes in people's diet structure, increased work and life pressure, reduced exercise, and deterioration of the living environment and other factors that are unfavorable to health. The incidence of various diseases, especially malignant diseases, is increasing year by year. Among them, liver cancer is one of the common malignant tumors in my country, with a high mortality rate. It ranks third in the order of malignant tumor deaths after stomach and esophagus, and ranks second in rural areas in some areas. Second only to gastric cancer, about 110,000 people die from liver cancer every year in my country, accounting for...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/53C12R1/84
Inventor 赵玉凤吴元欣雷明科邓凡池汝安黄锟赵锦
Owner WUHAN INSTITUTE OF TECHNOLOGY
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