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Subunit vaccine immunologic adjuvant and application thereof

A subunit vaccine and immune adjuvant technology is applied in the subunit vaccine immune adjuvant and application fields to achieve the effects of good effect, easy expression and enhanced immunogenicity

Inactive Publication Date: 2011-09-14
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are still some unsatisfactory aspects of genetically engineered vaccines in inducing the body's immune response. Therefore, researchers have tried to enhance the immune effect of fowl cholera subunit vaccines in many ways.

Method used

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  • Subunit vaccine immunologic adjuvant and application thereof
  • Subunit vaccine immunologic adjuvant and application thereof
  • Subunit vaccine immunologic adjuvant and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0030] The present invention will be further described below in conjunction with embodiment.

[0031] 1. Obtaining the complete nucleotide sequence of PG-1-GGGGS-3BS fusion peptide

[0032] The whole gene sequence of PG-1-GGGGS-3BS fusion peptide was amplified by overlapping extension PCR (SOE), and three primers were used to amplify. The three primers are F1 SEQ.ID.NO.3, F2 SEQ.ID.NO.4, F3 SEQ.ID.NO.5.

[0033] PCR reaction system 50 μl: 10×PCR Buffer 5 μL, dNTP (2.5 mM) 2 μL, primers F1, F2, F3 each 1 μL, rTeq DNA polymerase 0.5 μL, make up to 50 μL with deionized water.

[0034] PCR reaction program: pre-denaturation at 95°C for 5 min, amplification conditions: 94°C for 30 s, 57°C for 30 s, 72°C for 30 s, 35 cycles, and 72°C for 10 min. Nucleotide fragments with a size of 140 bp were recovered with a nucleic acid gel recovery kit produced by Dalian Takara Company. The electrophoresis diagram of its SOE-PCR product, the right side of the figure is the DL2000 nucleic acid mo...

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Abstract

The invention provides a subunit vaccine immunologic adjuvant and application thereof. The adjuvant is a fusogenic peptide of antibacterial peptide and allanoin tripeptide and has an amino acid sequence of SEQ.ID.NO.1 and a nucleotide sequence of SEQIDNO.2. In the invention, PG-1-GGGGS-3BS fusogenic peptide is used as a molecular adjuvant, and the immunogenicity of the outer membrane protein of chicken pasteurella multocida can be enhanced effectively; when the PG-1-GGGGS-3BS fusogenic peptide is used as a molecular adjuvant, a eukaryotic yeast expression vector is selected, the fusogenic peptide can be expressed and purified easily, can be modified by glycosylation and the like, and is easy for mass production; the fusogenic peptide expressed by yeast has modifier genes of eukaryotic cells, is similar to the structure of PG-1 expression in a mammal body and has good biological activity.

Description

[0001] technical field [0002] The invention relates to a technique for producing immune adjuvants by genetic engineering in the biotechnology pharmaceutical industry, in particular to a subunit vaccine immune adjuvant and its application. Background technique [0003] Antimicrobial peptides are the earliest immune active molecules produced by organisms in the long-term evolution process to adapt to the environment and survive. They have the characteristics of selective effects, small molecular weight, and no antigenicity. It plays an extremely important role in the immune defense against pathogenic invasion, so it is vividly called "natural antibacterial agent" or "cationic host defense peptide", which is an important molecular barrier for pathogenic invasion. Tani's research found that mouse splenocytes cultured in vitro can proliferate and increase cytokine production after being stimulated by human antimicrobial peptides, while in vivo treatment can lead to enhanced IgG...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61K39/102A61K39/108A61K39/112C07K19/00C12N15/62A61P31/04
Inventor 牛明福李翔宫强秦翠丽侯玉泽张敏王文辉吉萍
Owner HENAN UNIV OF SCI & TECH
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