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Polypeptide for preparing fishing immunologic adjuvant and purpose of polypeptide

An immune adjuvant, a technology for use, applied in the field of immunology, to achieve the effect of enhancing the efficacy of immune protection, simple preparation, and convenient use

Active Publication Date: 2016-04-13
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There is no report on the use of cytokines in the preparation of immune adjuvants for Streptococcus iniae vaccine

Method used

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  • Polypeptide for preparing fishing immunologic adjuvant and purpose of polypeptide
  • Polypeptide for preparing fishing immunologic adjuvant and purpose of polypeptide
  • Polypeptide for preparing fishing immunologic adjuvant and purpose of polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] The preparation of embodiment 1 polypeptide of the present invention

[0053] 1. Preparation of the gene of the present invention

[0054] Design primers: the 5' end of the upstream primer (P1) and the 5' end of the downstream primer (P2) plus enzyme cutting sites EcoRI and XhoI, the primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd., the sequence is as follows:

[0055] P15'-CG GAATTC GGAGAAGGAAAAGCAGAG-3' (the underlined part is the EcoRI restriction site);

[0056] P25'-CC CTCGAG TCAGTTTTGCTGTTTGATCT-3' (the underlined part is the XhoI restriction site).

[0057] Using channel catfish spleen cDNA as a template and P1 and P2 as primers, the IL-8 gene of the present invention (abbreviated as mCcIL-8) was amplified by PCR using a 25 μL reaction system, as shown in Table 1.

[0058] Table 1 The PCR amplification system of the gene of the present invention

[0059]

[0060] PCR reaction conditions: pre-denaturation at 95°C for 5 minutes; 30 c...

Embodiment 2

[0090] The preparation of embodiment 2 subunit vaccine of the present invention

[0091] 1. Antigen preparation

[0092] Using channel catfish spleen cDNA as template, P3 and P4 as primers,

[0093] P3: 5'-CGCGGATCCATGTCAATTATTACTGATG-3' (the underlined part is the BamHI restriction site);

[0094] P4: 5'-CGGAAGCTTCTATTTTTTTAGGTTGTAG-3' (the underlined part is the HindIII restriction site);

[0095] A 1308bp S.iniaeа-enolase gene was amplified;

[0096] The nucleotide sequence of the S. iniaeа-enolase gene is shown below:

[0097]

[0098] Insert the S.iniaeа-enolase gene into pET32a(+) to obtain pET32-eno, and further transfer it into BL21(DE3) competent cells to obtain recombinant strains;

[0099] The recombinant strain was induced with 0.1mMIPTG at 37°C for 4h, and the cells were collected by centrifugation. The supernatant was collected after the cells were ultrasonically disrupted, and purified according to the operating instructions of the Ni-NTA-SefinoseColumn ...

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Abstract

The invention discloses polypeptide shown in the amino acid sequence SEQ ID NO:1 and further discloses a purpose of polypeptide in preparing an immunologic adjuvant of a vaccine. The invention further discloses the immunologic adjuvant prepared from polypeptide and an adjuvant. By means of IL-8 polypeptide, the immunoprotection effect of the vaccine especially a streptococcus iniae subunit vaccine can be remarkable enhanced, and polypeptide can be used as the adjuvant of the vaccine. Polypeptide is safe to use, easy to prepare and convenient to use, and new effective choices are provided for prevention and treatment of fish diseases.

Description

technical field [0001] The invention belongs to the field of immunology, and in particular relates to a polypeptide for preparing fish immune adjuvant and its application. Background technique [0002] With the increasing degree of intensification of modern aquaculture, the losses caused by outbreaks of diseases in farmed fish are becoming more and more serious. Fish diseases have become an important factor hindering the healthy development of aquaculture. Streptococcus iniae (S. iniae) disease is a highly infectious bacterial disease that can cause explosive fish disease in a variety of wild or farmed fish, including freshwater and marine fish, resulting in Schools of fish die in large numbers. Moreover, Streptococcus iniae can also infect people through wounds, causing human cellulitis, endocarditis, arthritis, meningitis, and myelitis. [0003] Streptococcus dolphin disease is currently mainly controlled by chemical drugs, but chemotherapy has many disadvantages, which ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/54C12N15/24A61K39/39A61K39/09A61P31/04
CPCA61K39/092A61K39/39A61K2039/545A61K2039/55527C07K14/5421
Inventor 陈德芳汪开毓龙波王均王二龙耿毅
Owner SICHUAN AGRI UNIV
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