Actinobacillus pleuropneumoniae immune protective antigen protein APJL_0922 and application thereof

A protective antigen and actinobacillus technology, applied in the direction of bacterial antigen components, applications, bacterial peptides, etc., can solve the problems that limit the improvement and development of porcine pleuropneumonia subunit vaccines, and achieve strong immunogenicity and immune protection Effect

Active Publication Date: 2018-11-20
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 subun...

Method used

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  • Actinobacillus pleuropneumoniae immune protective antigen protein APJL_0922 and application thereof
  • Actinobacillus pleuropneumoniae immune protective antigen protein APJL_0922 and application thereof
  • Actinobacillus pleuropneumoniae immune protective antigen protein APJL_0922 and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1 Expression of APJL_0922 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 ce...

Embodiment 2

[0056] Example 2 Identification of the immune protection of APJL_0922 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_0922 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...

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Abstract

The invention discloses actinobacillus pleuropneumoniae immune protective antigen protein APJL_0922 and application thereof. The amino acid sequence of the actinobacillus pleuropneumoniae immune protective antigen protein is as shown in SEQ ID NO.1; and the immune protective antigen protein consists of 273 pieces of amino acid, and the mature polypeptide part is the 20th to the 273rd amino acid. The nucleotide sequence for encoding the immune protective antigen protein is preferably as shown in SEQ ID NO.2. The immune protective antigen protein has high immunogenicity and strong immunoprotection effect, and can be applied to preparation of a kit for detecting an actinobacillus pleuropneumoniae antibody, preparation of pig pleuropneumonia subunit vaccine and preparation of medicines for preventing diseases caused by the actinobacillus pleuropneumoniae. The invention provides a new material for preparing the pig pleuropneumonia subunit vaccine, and important significance in preventing and treating pig pleuropneumonia is achieved.

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_0922 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. Porcine pleuropneumonia is a highly contagious respiratory disease characterized by acute hemorrhagic, fibrinous, necrotizing bronchopneumonia and fibrinous pleurisy. Since the first report by Pattison et al. in 1957, the disease has spread to many countries around the world and regions, 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 immun...

Claims

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Application Information

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IPC IPC(8): C07K14/195C12N15/31C12N15/70G01N33/569A61K39/02A61P31/04
CPCA61K39/0208A61P31/04C07K14/195C12N15/70G01N33/56911
Inventor 刘金林祁超曹雨柔高露露张丽
Owner HUAZHONG NORMAL UNIV
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