Ab (Acinetobacter baumannii) Ata (acinetobacter trimeric autotransporter) protein immunogenicity

A technology of Acinetobacter baumannii and protein, which is applied in the direction of immunoglobulin, antibacterial drugs, recombinant DNA technology, etc., and can solve problems such as vaccine marketing

Active Publication Date: 2018-09-07
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Unfortunately, there is currently n...

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  • Ab (Acinetobacter baumannii) Ata (acinetobacter trimeric autotransporter) protein immunogenicity
  • Ab (Acinetobacter baumannii) Ata (acinetobacter trimeric autotransporter) protein immunogenicity
  • Ab (Acinetobacter baumannii) Ata (acinetobacter trimeric autotransporter) protein immunogenicity

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Experimental program
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Embodiment 1

[0040] Embodiment 1, the acquisition of fusion protein CTB-Ata

[0041] The fusion protein CTB-Ata includes 39 amino acids of the N-terminal α-helix part of CTB and the surface protein Ata (Acinetobacter trimeric autotransporter) of Acinetobacter baumannii.

[0042] 1. Fusion protein CTB-Ata

[0043] The surface protein Ata (Acinetobacter trimeric autotransporter) of Acinetobacter baumannii intercepts 39 amino acids of the N-terminal α-helix ( figure 1 ).

[0044] Then, the 39 amino acids of the α-helix part of the Ata protein, the tac promoter, and the CTB protein fused to form a fusion protein CTB-Ata are formed through the whole gene synthesis. The amino acid sequence of the fusion protein is sequence 2 in the sequence listing.

[0045] In the sequence 2, the 20th-123rd position is the fusion expressed CTB protein, the 127th-165th position is the α-helix part of the Ata protein, and the 201-206th position is the His tag.

[0046] The nucleotide sequence of the fusion pro...

Embodiment 2

[0057] Example 2, Functional Verification of Fusion Protein CTB-Ata

[0058] 1. Fusion protein animal immunization

[0059] The purified fusion protein CTB-Ata prepared in Example 1 was diluted with physiological saline to prepare an immunization sample containing the fusion protein CTB-Ata; immunization was carried out at an immunization dose of 40 μg fusion protein CTB-Ata per mouse.

[0060] Take 20 5-week-old (no significant difference in body weight) female Balb / c mice (Victoria Lihua), and randomly divide them into 2 groups (5 mice in each group):

[0061] Intraperitoneal immunization group: each mouse was injected with 100 μl of the immune sample containing the fusion protein CTB-Ata, and the immune dose was 40 μg per mouse;

[0062] Blank control group (no injection of any sample): female Balb / c mice;

[0063] Each group was immunized on the 1st, 14th, and 28th day respectively, and the blood was collected by docking the tail 10 days after each immunization injection...

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Abstract

The invention discloses Ab Ata protein immunogenicity and provides a protein which comprises Ab Ata protein N-terminal alpha-spiral part 39 amino acids and an adjuvant protein; the amino acid sequences of the Ab Ata protein N-terminal alpha-spiral part 39 amino acids are 127th-165th sites of the sequence 2 respectively. According to the Ab Ata, the N-terminal alpha-spiral part 39 amino acids are cut out, fused with CTB and expressed in BL21. Through ni-sepharose purification, peritoneal immunity is performed at 40 mu g/mice, the immunogenicity and the immune protective of the Ab Ata protein are verified through animal experiments, and the Ab Ata protein has excellent Ab infection resistance.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the immunogenicity of Ata protein of Acinetobacter baumannii. Background technique [0002] Acinetobacter baumannii (Ab) is a Gram-negative bacterium that widely exists in nature. One of the clinical pathogens. Widely distributed in hospital environments due to its rapid proliferation and strong adhesion, and the emergence of more and more resistant strains due to the heavy use of antibiotics, Acinetobacter baumannii has become a serious threat to healthcare systems worldwide. pose a serious threat. At present, antibiotics are mainly used clinically to combat the infection of Acinetobacter baumannii. However, the emergence of multi-drug resistant strains makes doctors have to combine drugs in treatment. However, there are many disadvantages in the combination drug, so the development of related vaccines is urgent to prevent the outbreak of Acinetobacter baumannii. Unf...

Claims

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Application Information

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IPC IPC(8): C07K14/21C07K19/00C12N15/31C12N15/62A61K39/104A61K39/39A61P31/04
CPCA61K39/1045A61K39/39A61P31/04C07K14/212A61K2039/55544C07K2319/55A61K2039/6037C07K16/1217
Inventor 朱力刘志成潘超王恒樑刘先凯王东澍冯尔玲
Owner ACADEMY OF MILITARY MEDICAL SCI
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