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Pleiomorphic saccharomyces hansenii expression recombination cholere toxin B subunit gene and its application

A technology of Hansenula polymorpha and cholera toxin, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as high cost, inability to industrialize, and pollution of CTA.

Inactive Publication Date: 2004-07-21
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are two methods to obtain CTB: one method is to isolate it from natural Vibrio cholerae, but this method has the risk of contaminating CTA and cannot be industrialized
Another method is to use genetically engineered bacteria to produce recombinant CTB (rCTB); the currently reported genetically engineered bacteria producing rCTB are only Escherichia coli, but the expression level is low and the cost is high

Method used

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  • Pleiomorphic saccharomyces hansenii expression recombination cholere toxin B subunit gene and its application
  • Pleiomorphic saccharomyces hansenii expression recombination cholere toxin B subunit gene and its application
  • Pleiomorphic saccharomyces hansenii expression recombination cholere toxin B subunit gene and its application

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Embodiment

[0023] 1. Establishment of codon usage of highly expressed genes in Hansenula yeast

[0024] Login to Genbank website http: / / www.ncbi.nlm.nih.gov The Hansenulaporlymorpha DNA sequence was retrieved, and the working draft and other non-essential sequences were excluded. After downloading to the local hard disk, use DNAStar software to remove the 5' and 3' untranslated regions of each sequence to obtain the coding sequence (CDS), and then use the DNAStar software MegAlign program to draw a phylogenetic tree with the ORF (open reading frame) of all genes, and save the file for hansenula.pau. then log in http: / / bioweb.pasteur.fr / seqanal / interfaces / codonw.html Website, enter the Codonw program entry, transfer the hansenula.pau file, and use the Codon W program to perform correspondence analysis (CA). The program automatically classifies the high-expression genes and low-expression genes of Hansenula, wherein the sample numbers of high-expression genes and low-expression genes e...

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Abstract

A polymorphous Hanson yeast expressed recombinant cholera toxin B subunit gene (CTB) is disclosed. In order to increase the expression of CTB in Hanson yeast, the coding gene of CTB is redesigned according to the application method of codon for the high-expression gene of Hanson yeast. The expression product can be used for preventing the diarrhea caused by cholera ribro and as the immune vaccine.

Description

technical field [0001] The invention belongs to the field of protein expression genetic engineering, and in particular relates to the use of a recombinant cholera toxin CTB subunit gene and the use of Hansenula polymorpha containing the gene as a cell factory to efficiently express CTB and the use of CTB and its fusion Application of protein in preparation of cholera vaccine and adjuvant. Background technique [0002] Cholera (Cholera) is one of the seven major epidemics in the world. It is designated as a Class A infectious disease in my country. It is an intestinal infectious disease caused by Vibrio cholerae (Vibrio cholerae), which causes acute diarrhea in humans and mammals during the onset , will soon die if not treated in time. Cholera toxin (CT) is a heat-labile enterotoxin secreted by Vibrio cholerae. The toxicity of CT is very strong, and humans are especially sensitive to CT, and 8 μg of CT can cause severe diarrhea. CT consists of two subunits, one A subunit (C...

Claims

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Application Information

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IPC IPC(8): A61K39/106A61K39/39C07K14/28C12N1/19C12N15/31C12N15/62C12N15/81C12P21/02
Inventor 邱并生宋厚辉李勇
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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