Riemerella anatipestifer DNA vaccine based on RA OmpA gene as well as preparation method and identification method of riemerella anatipestifer DNA vaccine
A technology of Duck Rimer's and DNA vaccines, applied in the biological field, can solve the problem of insufficient immune protection effect of the vaccine, and achieve strong immune protection effect and better immune protection effect
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Embodiment 1
[0043] A DNA vaccine based on RA OmpA gene Riemerella anatipestifer, including the eukaryotic expression plasmid pVAX1-OmpA of RA OmpA gene; the eukaryotic expression plasmid pVAX1-OmpA is extracted by Escherichia coli, and carried out with pVAX1 as a carrier Prepared, and each bottle of vaccine contains 100,000 μg of recombinant plasmid PVAX1-OmpA; when inoculated, the neck of ducklings is subcutaneously immunized with 100 μg of recombinant eukaryotic expression plasmid PVAX1-OmpA.
[0044] A preparation method based on RA OmpA gene Riemerella anatipestifer DNA vaccine as described above, comprising the following steps:
[0045] (1) Preparation of polyantiserum: RA OmpA gene was constructed into pET32a+ prokaryotic expression vector, and the recombinant expression plasmid pET32a+-RA-OmpA was transformed into competent cell BL21 to induce expression, and after IPTG induced protein expression, 12% SDS-PAGE After analyzing and confirming the target protein, put the recombinant p...
Embodiment 2
[0056] A DNA vaccine based on RA OmpA gene Riemerella anatipestifer, including the eukaryotic expression plasmid pVAX1-OmpA of RA OmpA gene; the eukaryotic expression plasmid pVAX1-OmpA is extracted by Escherichia coli, and carried out with pVAX1 as a carrier Prepared, and each bottle of vaccine contains 100,000 μg of recombinant plasmid PVAX1-OmpA; when inoculated, the neck of ducklings is subcutaneously immunized with 100 μg of recombinant eukaryotic expression plasmid PVAX1-OmpA.
[0057] The eukaryotic expression plasmid pVAX1-OmpA of the RA OmpA gene was identified by PCR, double enzyme digestion and sequencing. The results showed that the OmpA gene was correctly connected to the PVAX1 vector. The size of the OmpA gene fragment was about 1149bp. The double enzyme digestion and sequencing results were used to identify It is a positive recombinant eukaryotic expression plasmid, named PVAX1-OmpA, and the identification results are as follows: figure 1 , figure 2 shown.
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