Vaccine for preventing porcine epizootic diarrhea disease

A porcine epidemic diarrhea and vaccine technology, applied in the field of genetic engineering, can solve the problems of long antigen gene, difficult to guarantee safety, poor strain stability, etc., and achieve the effect of good stability, good protection and good safety

Inactive Publication Date: 2015-05-27
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, Xu Lili et al. constructed an attenuated Salmonella recombinant vaccine that can express the COE region and SD region of PEDV S protein. The antigen

Method used

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  • Vaccine for preventing porcine epizootic diarrhea disease
  • Vaccine for preventing porcine epizootic diarrhea disease
  • Vaccine for preventing porcine epizootic diarrhea disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1 Construction of recombinant bacteria of the present invention

[0052] 1 PEDV S 499-650 gene cloning

[0053] 1.1 Primer design

[0054] Referring to the S gene cDNA sequence of the PEDV SC-L strain published by GenBank (sequence number: KC886306), primers were designed using Primer 5.0 as follows (Table 1), wherein P1 / P2 were used to amplify S 499-650 Gene. Primers were synthesized by Dalian Bao Biological Engineering Co., Ltd.

[0055] Table 1 S 499-65 Gene amplification primer sequence and restriction site marking

[0056]

[0057] 1.2 PEDV S 499-650 gene amplification

[0058] 1.2.1 Extraction of PEDV RNA

[0059] Use the purified PEDV SC-L strain virus solution frozen in the laboratory as the material, and extract the viral RNA according to the instructions of the total RNA extraction kit. The operation method is as follows:

[0060] (1) Take 250 μL of virus liquid into a 2 mL EP tube, add 750 μL of RZ lysate, and beat several times with a ...

experiment example 1

[0175] Experimental example 1 Characteristic identification of recombinant attenuated Salmonella of the present invention

[0176] 1. Experimental method

[0177] Get the recombinant Salmonella SL7207 (pVAX-S) prepared in Example 1 499-650 ), according to the following method for identification:

[0178] 1 Biological characteristics of recombinant attenuated Salmonella

[0179] 1.1 Determination of growth curve of recombinant attenuated Salmonella

[0180] Pick SL7207 (pVAX-S 499-650 ) single colony, inoculated in 5mL LB liquid medium containing 100μg / mL Kan, until the bacterial solution OD 600 When the value is 0.6, inoculate 1 mL into 100 mL LB liquid medium containing 100 μg / mL Kan, culture at 37 °C, 100 r / min, for 24 h, absorb a small amount of bacterial liquid every 2 h, serially dilute and spread on three On the LB solid plate containing 100 μg / mL Kan, after culturing at 37°C for 18 hours, the colonies were counted, and the growth curves of different strains of reco...

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Abstract

The invention discloses an application of a recombinant salmonella containing a nucleotide sequence shown in SEQ ID NO:1 in preparation of a medicine for preventing a porcine epizootic diarrhea disease and also discloses a vaccine which is used for preventing the porcine epizootic diarrhea disease and is prepared from the recombinant salmonella containing the nucleotide sequence shown in SEQ ID NO:1 and pharmaceutically acceptable auxiliary materials or a carrier. The vaccine for preventing the porcine epizootic diarrhea disease can produce a high-level antibody specific to porcine epizootic diarrhea viruses, has a good protective effect on the porcine epizootic diarrhea disease and has a good clinical application prospect.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a vaccine for preventing porcine epidemic diarrhea disease. Background technique [0002] Porcine epidemic diarrhea virus is the causative agent of porcine epidemic diarrhea disease (Porcine epidemic diarrhea, PED). There are yellow curds. PEDV is mainly transmitted through the fecal-oral route. After the piglets in the farrowing house ingest the virus through the digestive tract, the virus mainly invades the epithelial cells of the small intestinal mucosa in the jejunum and ileum, and begins to replicate. The infection of PEDV leads to the decrease of the activity of disaccharidase (lactase, sucrase, maltase), aminopeptidase and alkaline phosphatase on the surface of epithelial cell membrane; at the same time, the normal structure of intestinal villi epithelial cells is destroyed and cell fragmentation occurs, resulting in intestinal The atrophy of the villi and the reductio...

Claims

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Application Information

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IPC IPC(8): A61K39/215A61K48/00A61P31/14A61P1/12C12N1/21C12R1/42
Inventor 文翼平黄小波梁恩涛文心田曹三杰伍锐陈杰张雨迪段素芬朱书权
Owner SICHUAN AGRI UNIV
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