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Immunoprotective antigen protein APJL_1976 for Actinobacillus pleuropneumoniae and application of immunoprotective antigen protein APJL_1976

A technology of protective antigen and actinobacillus, applied in the fields of application, vaccine, bacteria, etc., can solve the problems that limit the improvement and development of porcine pleuropneumonia subunit vaccine, and achieve the effect of strong immunogenicity and immune protection

Active Publication Date: 2018-11-16
HUAZHONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But at present, the number of immune protective antigen proteins that can be used to produce porcine pleuropneumonia subunit vaccines is relatively small, which largely limits the improvement and development of porcine pleuropneumonia subunit vaccines

Method used

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  • Immunoprotective antigen protein APJL_1976 for Actinobacillus pleuropneumoniae and application of immunoprotective antigen protein APJL_1976
  • Immunoprotective antigen protein APJL_1976 for Actinobacillus pleuropneumoniae and application of immunoprotective antigen protein APJL_1976
  • Immunoprotective antigen protein APJL_1976 for Actinobacillus pleuropneumoniae and application of immunoprotective antigen protein APJL_1976

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Expression of embodiment 1 APJL_1976 protein

[0024] 1. Genomic DNA extraction of Actinobacillus pleuropneumoniae

[0025] Take 1 mL of overnight cultured Actinobacillus pleuropneumoniae (JL03), centrifuge at 12,000 rpm for 1 minute, discard the supernatant, and then extract the genome of the JL03 strain according to the instructions of the genome extraction kit (Wuhan Boyue Biotechnology Co., Ltd.). The specific process is as follows: Add 200 μL RB to the bacterial pellet to resuspend, centrifuge at 10,000 rpm for 30 seconds, discard the supernatant, then add 200 μL RB to resuspend the pellet, then add 20 μL lysozyme to the centrifuge tube for vigorous shaking, and place at 37°C Incubate in the incubator for 15 minutes, then add 200 μL of binding solution CB, mix upside down, add 20 μL proteinase K (20 mg / mL), mix well, put it in a 70°C water bath for 10 minutes, take it out and add 100 μL iso Propanol, mix well by inversion, then transfer all the substances in the c...

Embodiment 2

[0056] Example 2 Identification of the immune protection of APJL_1976 protein

[0057] (1) Active immunization and challenge of mice

[0058] Six-week-old female BALB / c mice (purchased from the Experimental Animal Center of Hubei Provincial Center for Disease Control and Prevention) were immunized with APJL_1976 protein and GST protein purified in Example 1, 12 in each group. At the same time, a TSB blank control was set without immunization. The first immunization dose was 80 μg / mouse, mixed with an equal volume of Freund's complete adjuvant, emulsified, and injected subcutaneously at multiple points on the back of the mouse. After an interval of two weeks, the second immunization was carried out, the dose was 80 μg / mouse, mixed with an equal volume of Freund’s incomplete adjuvant, emulsified, and injected subcutaneously at multiple points on the back of the mouse. After 10 days, the antibody level was detected by ELISA, and HRP Labeled goat anti-mouse IgG was used as the s...

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Abstract

The invention discloses immunoprotective antigen protein APJL_1976 for Actinobacillus pleuropneumoniae and an application of the immunoprotective antigen protein APJL_1976. The amino acid sequence ofthe immunoprotective antigen protein for the Actinobacillus pleuropneumoniae is shown as SEQ ID NO.1, the immunoprotective antigen protein contains 273 amino acids, and a mature polypeptide part of the protein refers to 21-191-position amino acids. A nucleotide sequence for encoding the immunoprotective antigen protein is preferably shown as SEQ ID NO.2. The immunoprotective antigen protein has good immunogenicity and strong immunoprotective effect and can be applied to preparation of a kit for detecting an Actinobacillus pleuropneumoniae antibody, preparation of a subunit vaccine for porcinepleuropneumonia and preparation of a drug for preventing diseases caused by the Actinobacillus pleuropneumoniae. A new material is provided for preparing the subunit vaccine for porcine pleuropneumonia and has great significance in preventing and treating porcine pleuropneumonia.

Description

technical field [0001] The invention relates to the technical field of preparation of animal infectious disease subunit vaccines, in particular to an immunoprotective antigenic protein APJL_1976 of Actinobacillus pleuropneumoniae and its application. Background technique [0002] Actinobacillus pleuropneumoniae (Actinobacillus pleuropneumoniae) is a small Gram-negative bacillus, which is the pathogenic bacterium that causes swine pleuropneumonia. The disease is a highly contagious respiratory disease characterized by acute hemorrhagic, fibrinous, necrotizing bronchopneumonia and fibrinous pleurisy. Since Pattison et al. areas, seriously hindering the healthy development of the pig industry. [0003] Vaccine immunization is an effective means to prevent and control porcine pleuropneumonia. Porcine pleuropneumonia subunit vaccines are usually based on the identified immunoprotective antigen protein of Actinobacillus pleuropneumoniae, and are prepared through appropriate proc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/285C12N15/31C12N15/70C12N1/21G01N33/68A61K39/102A61P31/04C12R1/19
CPCA61K39/102A61K2039/552A61P31/04C07K14/285C12N15/70G01N33/6854
Inventor 祁超刘金林曹雨柔高露露张丽
Owner HUAZHONG NORMAL UNIV
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