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A method of improving fish anti-virus immunity

An anti-virus and immunity technology, applied in anti-viral agents, fish farming, pharmaceutical formulations, etc., can solve the problems of endangering the aquaculture industry, highly contagious fish viral diseases, and differences in serotypes, and achieve the goal of preventing and controlling fish. Virus-like diseases, the effect of enhancing anti-viral immunity

Active Publication Date: 2022-02-18
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Fish viral diseases are highly contagious, spread quickly, and have high mortality, seriously endangering the aquaculture industry and causing major economic losses
A small number of vaccines for fish viral diseases have been developed and produced at home and abroad, but there are still problems such as poor immune effect and serotype differences, which are difficult to apply in production practice

Method used

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  • A method of improving fish anti-virus immunity
  • A method of improving fish anti-virus immunity
  • A method of improving fish anti-virus immunity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: BMP8A gene knockout experiment

[0024] The zebrafish BMP8A protein DE amino acid sequence is SEQ ID NO: 1, and the amino acid sequence of the corresponding ripe peptide is SEQ ID NO: 2, the nucleotide sequence encoding the gene of the above BMP8A protein is SEQ ID NO: 3. The zebrafish BMP8A gene is mainly expressed in the essence of zebrafish, intestinal and skin tissue, the expression of the heart, the heart, muscle tissue is relatively low in brain tissue ( figure 1 .

[0025] Using TALEN technology to knock out of the zebrafish BMP8A gene. The specific method is as follows:

[0026] 1) Target site design:

[0027] Using Submit Talen Targeter (https: / / tale-nt.cac.cornell.edu / node / add / Talen) website, according to the TALEN technology target site design principle, each exon of the zebrafish BMP8A gene is analyzed, combined ZBMP8A gene domain features, design target sites, and find high efficiency enzyme sinks through the NEB website. The target site is selected ...

Embodiment 2

[0044] Example 2: GcrV virus stimulation can significantly increase the expression of zebrafish BMP8A gene

[0045] Zebrafish in the abdominal cavity of grass carp reovirus, GCRV was used to detect the expression of BMP8A gene in spleen, intestinal, liver and kidney tissue, and found that GCRV virus stimulation can significantly raise zebra. Fish BMP8A gene expression ( Figure 7 . Similarly, the zebrafish liver dirty cell line (ZFL) was treated with GCRV virus, and also found that the expression of BMP8A gene was significantly up-regulated. Figure 8 . The results show that the zebrafish BMP8A has a significant response to virus stimulation, and BMP8A has an important role in fish antiviral immunization.

Embodiment 3

[0046] Example 3: Overgraded zebrafish BMP8A can significantly adjust the expression of virus-related genes

[0047]In the zebrafish ZFL cell line, the expression of zebrafish BMP8A was expressed, and the expression of antiviral-related gene IFN φ1, IFNφ3 and MX was detected by Real-Time PCR technology. It was found that the expression of zebrafish BMP8A can significantly adjust the virus-related gene IFN φ1, IFNφ3 and MX expression ( Figure 9 ) Further proves that BMP8A has an important role in fish antiviral immunization.

[0048] In the EPC cell line, zebrafish BMP8A was expressed. Cells were infected with GcrV virus, and the expression of viral gene vp5 was detected by Real-Time PCR technology. It was found that the expression of zebrafish BMP8A can significantly decrease the expression of viral gene vp5 ( Figure 10 It is proved that BMP8A has a function of suppressing replication in viral in EPC cells.

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Abstract

The invention provides a method for improving the antiviral immunity of fish, which enhances the antiviral immunity of fish by up-regulating the expression of Bmp8a protein. The present invention first provides an application of the Bmp8a protein, which is the application in the preparation of a product for improving the antiviral immunity of fish. The present invention proves that the antiviral ability of zebrafish after the knockout of bmp8a gene is significantly reduced, while overexpression of Bmp8a can significantly upregulate the expression of antiviral related genes such as interferon, showing that Bmp8a plays an important role in antiviral immunity. By injecting Bmp8a mature peptide into fish, the antiviral immunity of fish is enhanced, so as to achieve the purpose of preventing and treating viral diseases of fish.

Description

Technical field [0001] The present invention belongs to the field of fish immunization, and more particularly to a method of improving fish anti-viral immunity. Background technique [0002] Fish viral disease is strong, fast communication speed, high mortality, serious harm to aquaculture industry, can cause significant economic losses. At home and abroad have been developed and produced a vaccine for a few fish viral diseases, but there is still a problem of poor immunity, and the variation of serotype is difficult to apply in production practice. Therefore, there is still a considerable difficulty in the treatment of fish viral diseases, mainly based on prevention. [0003] Bone Morphogenetic Protein (BMP) family is a kind of hydrophobic glycoprotein family, belonging to Transforming Growth Factor Beta (TGF-β) superfoot. BMP8 is one of the many members of the BMP family. In zebrafish, BMP8 is only a form of BMP8A. The zebrafish BMP8A first formed a pre-peptide containing 433 a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/18A61P31/12A61P37/04A23K50/80A23K20/147A01K61/13G01N27/447
CPCA61K38/1841A61K9/0019A61P37/04A61P31/12A01K61/13A23K50/80A23K20/147G01N27/447Y02A40/81
Inventor 刘振辉钟慎杰王颖张士璀
Owner OCEAN UNIV OF CHINA
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