Immunoprotective antigen protein APJL_1380 for actinobacillus pleuropneumoniae and application thereof

A technology of protective antigen and actinobacillus, applied in the direction of bacterial antigen composition, application, bacteria, etc., can solve the problems that limit the improvement and development of porcine pleuropneumonia subunit vaccines, and achieve strong immunogenicity and immune protection effects

Active Publication Date: 2018-11-13
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

Method used

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  • Immunoprotective antigen protein APJL_1380 for actinobacillus pleuropneumoniae and application thereof
  • Immunoprotective antigen protein APJL_1380 for actinobacillus pleuropneumoniae and application thereof
  • Immunoprotective antigen protein APJL_1380 for actinobacillus pleuropneumoniae and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0023] The expression of embodiment 1APJL_1380 protein

[0024] 1. Extraction of genomic DNA of Actinobacillus pleuropneumoniae

[0025] Take 1 mL of the bacterial liquid of Actinobacillus pleuropneumoniae (JL03) cultured overnight, 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 of RB to the cell pellet to resuspend, centrifuge at 10,000 rpm for 30 seconds, discard the supernatant, add 200 μL of RB to resuspend the pellet, and then add 20 μL of lysozyme to the centrifuge tube, shake vigorously, and place it at 37°C Incubate in an incubator for 15 minutes, then add 200 μL of binding solution CB, invert and mix, add 20 μL of proteinase K (20 mg / mL), mix well, put it in a 70°C water bath for 10 minutes, take it out and cool it and add 100 μL of isozyme. Propanol, invert and...

Embodiment 2

[0056] The identification of embodiment 2APJL_1380 protein immunoprotective

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

[0058] 6-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_1380 protein and GST protein purified in Example 1, 12 mice in each group. At the same time, a blank control of TSB was set, and no immunization was performed. The first immunization dose was 80 μg / mouse, mixed with an equal volume of Freund's complete adjuvant and emulsified, and inoculated at multiple points subcutaneously on the back of mice. After an interval of two weeks, the second immunization was carried out at a dose of 80 μg / mice, mixed with an equal volume of incomplete Freund's adjuvant and emulsified, and subcutaneously injected into the back of the mice at multiple points. The labeled goat anti-mouse IgG was the secondary antibody (1:5000 dilution). The titer o...

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Abstract

The invention discloses immunoprotective antigen protein APJL_1380 for actinobacillus pleuropneumoniae and application thereof. An amino acid sequence of the immunoprotective antigen protein for the actinobacillus pleuropneumoniae is shown in SEQ ID NO.1. The immunoprotective antigen protein consists of 273 amino acids, and the matured polypeptide part has amino acids at the 25th to 582nd sites; anucleotide sequence for coding the immunoprotective antigen protein is shown in SEQ ID NO.2. The immunoprotective antigen protein has the advantages that the immunogenicity is good, and the immunoprotective function is strong; the immunoprotective antigen protein can be applied to preparation of kits for detecting actinobacillus pleuropneumoniae antibodies, preparation of swine pleuropneumoniae sub-unit vaccines, and preparation of medicines for preventing diseases caused by the actinobacillus pleuropneumoniae; a novel material is provided for the preparation of the swine pleuropneumoniae sub-unit vaccines, and the important meaning is realized in the preventing and treatment of the swine pleuropneumoniae.

Description

technical field [0001] The invention relates to the technical field of animal infectious disease subunit vaccine preparation, in particular to an immunoprotective antigen protein APJL_1380 of Actinobacillus pleuropneumoniae and its application. Background technique [0002] Actinobacillus pleuropneumoniae (Actinobacillus pleuropneumoniae) is a small gram-negative bacterium that causes porcine pleuropneumoniae. The disease is a highly contagious respiratory disease characterized by acute hemorrhagic, fibrinous, necrotizing bronchopneumonia and fibrinous pleurisy. Since it was first reported by Pattison et al in 1957, the disease has spread to many countries and regions around the world. region, seriously hindering the healthy development of the pig industry. [0003] Vaccine immunization is an effective means to prevent and control porcine pleuropneumonia. Porcine pleuropneumoniae subunit vaccines are usually prepared by appropriate processing based on the identified immuno...

Claims

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Application Information

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IPC IPC(8): C07K14/195C12N15/31C12N15/70C12N1/21G01N33/569A61K39/02A61P31/04C12R1/19
CPCA61K39/02A61P31/04C07K14/195C12N15/70G01N33/56911
Inventor 刘金林祁超曹雨柔高露露张丽
Owner HUAZHONG NORMAL UNIV
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