Preparation method and application of rhabdovirus expressed swine fever E2 subunit vaccine
A subunit vaccine and baculovirus technology, which is applied in the field of preparation of swine fever E2 subunit vaccine, can solve the problems of not being able to prevent and control swine fever, and can not distinguish between infected and immunized animals, and achieve good safety and immunogenicity Strong, high-yielding effect
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[0049] Example 1 Expression of Classical Swine Fever SP-E2 Protein
[0050] 1) Amplification of the target gene
[0051] According to Genbank design primers SP-F, SP-R, E2-F, E2-R, first amplified with SP-F, SP-R to obtain the SP signal peptide, and amplified with E2-F, E2-R to obtain the E2 fragment. Then SP and E2 are used as templates, and SP-F and E2-R primers are amplified to obtain the target gene SP-E2 (such as figure 2 Shown).
[0052] 2) Construction of shuttle plasmid
[0053] The pFastBacHTB plasmid was digested with EcoRI and XholⅠ restriction enzymes at 37°C for 2 hours and recovered. The digested vector and the target gene were recombined with homologous recombinase for 30 minutes at 37°C, transformed into Ecoli competent cells, spotted and identified, and positive bacteria were sent Sequencing.
[0054] 3) Acquisition of recombinant bacmid
[0055] Transform the positive plasmid into DH10Bac competent cells, select leukoplakia 48h later, and perform PCR identification w...
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[0066] Example 2 Preparation and animal immunization experiment of swine fever SP-E2 protein subunit vaccine of the present invention.
[0067] The SP-E2 protein expressed in Example 1 and Seppic 206 water adjuvant were mixed at a ratio of 1:1 at 350 rpm for 10 minutes, and stored at 4°C and protected from light.
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