Application of il-17a protein in preparation of animal vaccine adjuvant

A 1. IL-17A, protein technology, applied in the field of aquaculture, can solve problems such as weak immune response

Active Publication Date: 2020-07-07
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the immune response elicited by oral antigen delivery alone is often weak

Method used

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  • Application of il-17a protein in preparation of animal vaccine adjuvant
  • Application of il-17a protein in preparation of animal vaccine adjuvant
  • Application of il-17a protein in preparation of animal vaccine adjuvant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1, preparation of adjuvant and identification of its biological activity

[0057] 1. Cloning of rainbow trout IL-17A gene

[0058] According to the nucleotide sequence of rainbow trout IL-17A gene (GenBank: NM_001124619), the primer F1: 5'- GGATCC AAAGGAATGAAGGTGACAAAGG-3' (single underline is the recognition site of restriction endonuclease BamHI), primer R1: 5'- AAGCTT TTATCAAGTAGTCCTTGCCCA-3' (single underline is the recognition site of restriction endonuclease HindⅢ), primer F2: 5'-GAATTC ATGAAAGGAATGAAGGTGACAAAGG-3' (the single underline is the recognition site of the restriction endonuclease EcoRI, and the double underline is the kozak sequence) and primer R2: 5'- CTCGAG TCAAGTAGTCCTTGCCCA-3' (single underline is the recognition site of restriction endonuclease XhoI).

[0059] 1. Execute the rainbow trout, quickly remove the head kidney and spleen aseptically on ice, and then add an appropriate amount of MEM medium containing 10% (v / v) FBS. After...

Embodiment 2

[0118] The application of the adjuvant prepared in embodiment 2, embodiment 1

[0119] 1. Preparation of DNA vaccine (recombinant plasmid pEE12.4-G)

[0120] 1. Artificially synthesize the double-stranded DNA molecule shown in sequence 5 in the sequence listing and use it as a template, using primer pair C (by primer F3: 5'- GAATTC ATGGACACCATGATCACCACTCCG-3' (single underline is the recognition site of restriction endonuclease EcoRI) and primer R3: 5'- GGATCC TCAGGACCGGTTTGCCAGGTGAT-3' (single underline is the recognition site of the restriction endonuclease BamHI) to perform PCR amplification to obtain the PCR amplification product C of 1539bp.

[0121] 2. Take the PCR amplification product C, perform double digestion with restriction endonucleases EcoRI and BamHI, and recover the digested product C of about 1539bp.

[0122] 3. Take the pEE12.4 vector, perform double digestion with restriction endonucleases EcoRI and BamHI, and recover about 6.9kb of vector backbone C. ...

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Abstract

The invention discloses application of IL-17A protein in preparing animal vaccine adjuvants. The IL-17A protein is protein shown as an amino acid sequence 2 in a sequence table. Experiments prove that compared with rainbow trout injected with infectious hematopoietic necrosis virus G protein, expression level of IgT gene, IgM gene, Mx gene, Viperin gene and CD8 gene of the rainbow trout injected with the infectious hematopoietic necrosis virus G protein and the IL-17A protein is improved remarkably, valence of neutralizing antibody is improved remarkably, proliferation of lymphocytes is remarkably enhanced, and protection force of the infectious hematopoietic necrosis virus G protein is effectively increased. Therefore, the IL-17A protein has important application value in preparing the animal vaccine adjuvants.

Description

technical field [0001] The invention relates to the field of breeding, in particular to the application of IL-17A protein in the preparation of animal vaccine adjuvants. Background technique [0002] Fish and aquatic products are still an important source of food and nutrition for humans all over the world. At present, rainbow trout is an important component of commercial cold-water fish in most countries. However, with the increase of breeding density and the deterioration of the breeding environment, rainbow trout farming is often threatened by infectious diseases. Vaccine has been proved to be an effective means to prevent and control pathogenic microorganisms in rainbow trout. [0003] Many countries restrict the use of live vaccines due to biosafety concerns, while inactivated or genetically engineered vaccines do not provide the expected protective effect in some cases. In addition, injection immunization is currently the main method of fish vaccination, which is not...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39A61K39/205A61K48/00C12N15/24C07K14/54A61P31/14
CPCA61K39/12A61K39/39A61K2039/55527C07K14/5418C12N2760/20034
Inventor 曹永生卢彤岩徐黎明李绍戊王荻赵景壮刘红柏
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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