Tuberculosis candidate vaccine fusion protein

A technology of recombinant protein and Mycobacterium tuberculosis, applied in the field of fusion protein, can solve the problems of unstable immune protection effect and great difference in immune protection effect, and achieve the effect of excellent immunogenicity

Active Publication Date: 2020-08-11
SICHUAN UNIV
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  • Abstract
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Problems solved by technology

BCG (Bacille Calmette Guerin, BCG) is currently the only vaccine used to prevent tuberculosis, but its immune protection effect is extremely unstable, and the immune protection effect of peop

Method used

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  • Tuberculosis candidate vaccine fusion protein
  • Tuberculosis candidate vaccine fusion protein
  • Tuberculosis candidate vaccine fusion protein

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Experimental program
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Effect test

Embodiment Construction

[0019] 1. Experimental main reagents

[0020] Medium reagents: tryptone (Tryptone) and yeast extract (Yeast Extract) were purchased from Oxoid, UK;

[0021] Gene cloning reagents: NdeI, Hind III restriction endonuclease, and T4 DNA ligase were purchased from Thermo Company, Germany; dNTP and rTaq enzymes were purchased from Takara Company; E.Z.N.A. Plasmid Mini Kit I, E.Z.N.A. GelExtraction Kits were purchased from Omega Company in the United States; PCR primers were synthesized by Shanghai Sangon Biotechnology Company; pet28a vector and H37Rv strain genome were preserved in our laboratory; BL21 (DE3) competent cells were purchased from Shanghai Sangon Biotechnology Company; RPMI1640 medium and Fetal bovine serum was purchased from Gibco. Esat6 monoclonal antibody was purchased from Santa Cruz Company of the United States; cytokine ELISA kit was purchased from Boxinsheng Company. The endotoxin removal kit ToxinEraser was purchased from GenScript;

[0022] Antibiotics: Ka...

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Abstract

The invention relates to construction of a chimeric protein Dodecin-ESAT6 candidate vaccine of an ESAT6 gene and an RV1498a gene of mycobacterium tuberculosis, and immunogenicity research of the chimeric protein Dodecin-ESAT6 candidate vaccine, i.e., through a genetic engineering technique, sequences of the ESAT6 gene and the RV1498a gene of the mycobacterium tuberculosis are inserted into sequences of the same escherichia coli plasmid pET28a, and a recombinant plasmid rpET28a:RV1498a-ESAT6 is constructed; and by a heat shock transforming method, carriers are guided into escherichia coli, andfusion protein Dodecin-ESAT6 is expressed. The expressed fusion protein Dodecin-ESAT6 is constructed, and the immunogenicity is better than that of ESAT6. The invention provides a preparation processof the fusion protein Dodecin-ESAT6, researches the immunogenicity of the fusion protein Dodecin-ESAT6, and belongs to the field of genetic engineering and the field of tuberculosis vaccines. Occurrence and propagation of tuberculosis can be effectively prevented.

Description

Technical field: [0001] The invention relates to the fields of genetic engineering and novel tuberculosis vaccines, in particular to a fusion protein. Background technique: [0002] Tuberculosis (tuberculosis, TB) is a chronic infectious disease that seriously endangers human health. It is caused by mycobacteria (mainly Mycobacterium tuberculosis, also known as "tuberclebacillus"). In recent years, population flow, Human immunodeficiency virus (HIV) and Mycobacterium tuberculosis co-infection, Mycobacterium tuberculosis multidrug resistance, and other factors have brought new challenges to the prevention and treatment of tuberculosis. BCG (Bacille Calmette Guerin, BCG) is currently the only vaccine used to prevent tuberculosis, However, its immune protection effect is extremely unstable, and the immune protection effect of populations in different regions varies greatly after inoculation with BCG (the protection rate ranges from 0% to 80%). It is urgent to develop a tubercul...

Claims

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Application Information

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IPC IPC(8): C07K14/35C12N15/70A61K39/04A61P31/06
CPCC07K14/35C12N15/70A61K39/04A61P31/06A61K2039/523
Inventor 赵容川鲍朗
Owner SICHUAN UNIV
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