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
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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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