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Method for improving antiviral immunity of fishes

An anti-virus and immunity technology, applied in anti-viral agents, fish farming, pharmaceutical formulations, etc., can solve problems such as poor immune effect, harm to aquaculture, and high infectivity of fish viral diseases.

Active Publication Date: 2019-08-13
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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  • Method for improving antiviral immunity of fishes
  • Method for improving antiviral immunity of fishes
  • Method for improving antiviral immunity of fishes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: bmp8a gene knockout experiment

[0024] The DE amino acid sequence of the zebrafish Bmp8a protein is SEQ ID NO:1, the amino acid sequence of its corresponding mature peptide is SEQ ID NO:2, and the nucleotide sequence of the gene encoding the above-mentioned Bmp8a protein is SEQ ID NO:3. The zebrafish bmp8a gene is mainly expressed in the zebrafish testis, intestine and skin tissue, followed by the expression in the brain tissue, and the expression level in the heart and muscle tissue is relatively low ( figure 1 ).

[0025] The zebrafish bmp8a gene was knocked out using TALEN technology. The specific method is as follows:

[0026] 1) Target site design:

[0027] Use the Submit TALEN Targeter (https: / / tale-nt.cac.cornell.edu / node / add / talen) website to analyze each exon of the zebrafish BMP8a gene according to the design principles of TALEN technology target sites, and combine According to the characteristics of the zBMP8a gene structure domain, design ...

Embodiment 2

[0044] Example 2: GCRV virus stimulation can significantly up-regulate the expression of zebrafish bmp8a gene

[0045] Zebrafish were injected intraperitoneally with Grass carp reovirus (GCRV), and the effects on bmp8a gene expression in spleen, intestine, liver and kidney tissues were detected by Real-time PCR technology, and it was found that GCRV virus stimulation could significantly up-regulate the zebrafish Expression of fish bmp8a gene ( Figure 7 ). Similarly, the expression of bmp8a gene was also found to be significantly up-regulated when the zebrafish liver cell line (ZFL) was treated with GCRV virus ( Figure 8 ). The results showed that zebrafish bmp8a had an obvious response to virus stimulation, and Bmp8a played an important role in fish antiviral immunity.

Embodiment 3

[0046] Example 3: Overexpression of zebrafish Bmp8a can significantly up-regulate the expression of anti-virus-related genes

[0047]In the zebrafish ZFL cell line, zebrafish Bmp8a was overexpressed, and the expression of anti-virus related genes IFNφ1, IFNφ3 and Mx was detected by Real-time PCR technology, and it was found that overexpression of zebrafish Bmp8a could significantly up-regulate the anti-virus related genes Expression of Mx ( Figure 9 ), further proving that Bmp8a plays an important role in fish antiviral immunity.

[0048] In the EPC cell line, zebrafish Bmp8a was overexpressed. The cells were infected with GCRV virus, and the expression of the viral gene VP5 was detected by Real-time PCR technology. It was found that overexpression of zebrafish Bmp8a could significantly down-regulate the expression of the viral gene VP5 ( Figure 10 ), proving that Bmp8a has the function of inhibiting virus replication in EPC cells.

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Abstract

The invention provides a method for improving the antiviral immunity of fishes. The antiviral immunity of the fishes is enhanced by increasing the expression amount of Bmp8a protein. The invention firstly provides use of the Bmp8a protein, which is application of the Bmp8a protein in the preparation of a product for improving the antiviral immunity of the fishes. According to the method, it is proved that the antiviral capacity of zebra fishes after bmp8a gene knockout is significantly reduced, over-expression of the Bmp8a can significantly up-regulate the expression of antiviral related genessuch as interferon, and it is shown that the Bmp8a plays an important role in the antiviral immunity. Bmp8a mature peptides are injected in the fishes, which enhances the antiviral immunity of the fishes, thereby achieving the purpose of preventing and treating viral diseases of the fishes.

Description

technical field [0001] The invention belongs to the technical field of fish immunity, and in particular relates to a method for improving fish anti-virus immunity. Background technique [0002] Fish viral diseases are highly contagious, spread quickly, and have high mortality rates, which seriously endanger the aquaculture industry and can cause 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. Therefore, there are still considerable difficulties in the treatment of fish viral diseases at present, and the main focus is on prevention. [0003] Bone Morphogenetic Protein (BMP) family is a family of hydrophobic glycoproteins, belonging to the Transforming Growth Factor Beta (TGF-β) superfamily. BMP8 is one of many members of the BMP family, and in zebrafish, Bmp8 has ...

Claims

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Application Information

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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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