Nucleic acid molecule and vaccine for preventing or treating feline infectious peritonitis as well as preparation method and application of nucleic acid molecule and vaccine
By developing specific nucleic acid fragments and recombinant vector technologies, the problem of unsatisfactory effect of cat abdominal virus vaccine in the prior art has been solved, efficient and convenient vaccine production and use have been achieved, significantly improving the cure rate of cat abdominal disease and reducing treatment costs.
Patent Information
- Application Number
- CN202510294847.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Priority Date
- 2024-05-17
- Filing Date
- 2025-03-13
- Publication Date
- 2025-06-06
AI Technical Summary
The prior art is difficult to effectively prevent or treat infectious peritonitis in cats, especially due to the unsatisfactory vaccine results from antibody-dependent enhancement effect (ADE).
A new ultra-low-cost, efficient therapeutic vaccine preparation for feline abdominal virus was developed. By isolating specific nucleic acid fragments such as S1-N, S1-C, S2, E gene, N gene, etc., expressing polypeptides or proteins as immunogens, and is used to induce an immune response against feline abdominal virus, and is achieved through recombinant vector and bacterial vector vaccine technology.
It has achieved the improvement of the cure rate of cat abdominal disease in a shorter dosage cycle, reduced the occurrence of ADE, and achieved ultra-low-cost vaccine production through improved engineered bacteria as production carriers, and it is more convenient to use oral capsule preparations.
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Figure CN120099039A_ABST
Abstract
Claims
1. An isolated nucleic acid molecule, comprising nucleic acid fragments S1-N, S1-C, S2, E gene, N gene, S1-N (UU2), S1-C (UU2) and S2 (UU2); the protein expressed by the nucleic acid fragments is used as an immunogen to induce the production of an anti-feline infectious peritonitis virus therapeutic vaccine; The nucleotide sequence of the nucleic acid fragment N gene is shown in SEQ ID NO: 1; The nucleotide sequence of the nucleic acid fragment E gene is shown in SEQ ID NO: 2; The nucleotide sequence of the nucleic acid fragment S1-N is shown in SEQ ID NO: 3; The nucleotide sequence of the nucleic acid fragment S1-C is shown in SEQ ID NO: 4; The nucleotide sequence of the nucleic acid fragment S2 is shown in SEQ ID NO: 5; The nucleotide sequence of the nucleic acid fragment S1-N(UU2) is shown in SEQ ID NO: 6; The nucleotide sequence of the nucleic acid fragment S1-C(UU2) is shown in SEQ ID NO: 7; The nucleotide sequence of the nucleic acid fragment S2 (UU2) is shown in SEQ ID NO:
8.
2. An isolated nucleic acid molecule according to claim 1, characterized in that The nucleic acid fragments S1-N, S1-C, S2, E gene, N gene, S1-N (UU2), S1-C (UU2) and S2 (UU2) are designed with feline coronavirus structural protein Spike (S), N gene and E gene as screening target antigens.
3. An isolated nucleic acid molecule according to claim 2, characterized in that The method for designing the nucleic acid fragment comprises the following steps: S1: Feline coronavirus structural protein Spike (S), N gene, and E gene were used as target antigens for screening; S2: The functional domains of multiple target proteins were analyzed, and eight target proteins or target protein truncations, namely S1-N, S1-C, S2, E gene, N gene, S1-N (UU2), S1-C (UU2) and S2 (UU2), were designed; the sequence of the target protein was optimized based on the Escherichia coli expression system; and the nucleic acid fragments described in SEQ ID NO: 1-8 were obtained. A recombinant vector comprising the nucleic acid molecule according to any one of claims 1 to 3.
5. The method for constructing a recombinant vector according to claim 4, characterized in that: Using pBAD-his as an expression plasmid vector, inserting any nucleic acid fragment described in claim 1 into the expression plasmid vector, designing and constructing a pBAD-ClyA-Ags recombinant vector.
6. A vaccine for preventing or treating feline infectious peritonitis, comprising the nucleic acid molecule according to any one of claims 1 to 3 or the recombinant vector according to claim 4.
7. A vaccine for preventing or treating feline infectious peritonitis as claimed in claim 6, characterized in that The vaccine comprises a bacterial vector vaccine, arabinose and a pharmaceutically acceptable carrier.
8. A vaccine for preventing or treating feline infectious peritonitis as claimed in claim 7, characterized in that The bacterial vector used in bacterial vector vaccines is Gram-negative bacteria, and this patent uses Escherichia coli.
9. A vaccine for preventing or treating feline infectious peritonitis as claimed in claim 8, characterized in that The method for preparing a bacterial vector vaccine is as follows: the recombinant vector pBAD-ClyA-Ags described in claim 5 is introduced into Escherichia coli respectively, and the bacteria are cultured to the logarithmic growth phase to prepare a bacterial vaccine; the recombinant vector pBAD-ClyA-Ags has an arabinose-induced araBAD promoter, which can induce the expression of the target antigen at a concentration of 10g / L-20g / L arabinose.
10. A vaccine for preventing or treating feline infectious peritonitis as claimed in claim 9, characterized in that: The Gram-negative bacteria can secrete outer membrane vesicles, and the outer membrane vesicles can efficiently deliver target antigens, activate the immune response of the body, and exert antiviral immune effects.
11. A vaccine for preventing or treating feline infectious peritonitis as claimed in claim 9, characterized in that: The bacterial vaccine is loaded in a capsule, and the capsule is used for oral administration.