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Application of IL-17A protein in preparing animal vaccine adjuvants

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

Active Publication Date: 2017-11-21
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 preparing animal vaccine adjuvants
  • Application of IL-17A protein in preparing animal vaccine adjuvants
  • Application of IL-17A protein in preparing animal vaccine adjuvants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 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'- was designed and artificially synthesized. GGATCC AAAGGAATGAAGGTGACAAAGG-3' (single underline is the recognition site of restriction enzyme BamHI), primer R1: 5'- AAGCTT TTATCAAGTAGTCCTTGCCCA-3' (single underline is the recognition site of restriction endonuclease HindⅢ), primer F2: 5'-GAATTC ATGAAAGGAATGAAGGTGACAAAGG-3' (single underline is the recognition site of restriction enzyme EcoRI, double underline is kozak sequence) and primer R2: 5'- CTCGAG TCAAGTAGTCCTTGCCCA-3' (single underline is the recognition site of restriction enzyme XhoI).

[0059] 1. The rainbow trout was killed, the kidney and spleen were quickly aseptically peeled off on ice, and then an appropriate amount of MEM medium containing 10% (v / v) FBS was added. A...

Embodiment 2

[0118] Example 2. Application of the adjuvant prepared in Example 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 table and use it as a template, using primer pair C (from primer F3: 5’- GAATTC ATGGACACCATGATCACCACTCCG-3' (single underline is the recognition site of restriction enzyme EcoRI) and primer R3: 5'- GGATCC TCAGGACCGGTTTGCCAGGTGAT-3' (single underline is the recognition site of restriction endonuclease BamHI) was subjected to PCR amplification to obtain PCR amplification product C of 1539 bp.

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

[0122] 3. Take the pEE12.4 vector and double-enzyme digestion with restriction enzymes EcoRI and BamHI to recover the vector backbone C of about 6.9kb.

[0123] 4. Connect the digested produc...

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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 important sources 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, but with the increase in breeding density and the deterioration of the breeding environment, rainbow trout farming is often threatened by infectious diseases. Vaccines have been proved to be an effective means to prevent and control the pathogenic microorganisms of rainbow trout. [0003] Many countries restrict the use of live vaccines due to biosecurity concerns, but inactivated vaccines or genetically engineered vaccines cannot provide the expected protective effects in some cases. In addition, injection immunization is the main method of fish vaccination. This method is not o...

Claims

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Application Information

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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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