Swine composite interferon used for emergent prevention for African swine fever

[0034] Embodiment 1, the preparation of protein solution

[0035] 1. Preparation of recombinant plasmids

[0036] The double-stranded DNA molecule shown in nucleotides 4-435 of Sequence 2 in the sequence listing was inserted between the NdeI and XhoI restriction sites of the pET30a(+) vector to obtain recombinant plasmid I. The recombinant plasmid I has an open reading frame shown in sequence 2 of the sequence listing, and expresses the protein shown in sequence 1 of the sequence listing.

[0037] The double-stranded DNA molecule shown in nucleotides 4-948 of Sequence 4 in the sequence listing was inserted between the NdeI and XhoI restriction sites of the pET30a(+) vector to obtain recombinant plasmid II. The recombinant plasmid II has an open reading frame shown in sequence 4 of the sequence listing, and expresses the protein shown in sequence 3 of the sequence listing.

[0038] Insert the double-stranded DNA molecule shown in nucleotides 4-552 of Sequence 6 in the sequence l

[0080] Embodiment 2, detect interferon activity

[0081] The test solutions were: the protein I solution, protein II solution, protein III solution or protein IV solution prepared in Example 1, all of which were freshly prepared.

[0082] 1. Cell plating

[0083] Get well-grown PK-15 cells, digest with trypsin, and then suspend with DMEM medium containing 100 U / mL penicillin, 100 U / mL streptomycin and 10% (volume ratio) FBS to obtain a cell concentration of 1 × 10 6 cell suspension per mL, add the cell suspension to a 96-well cell culture plate (100 μl / well), and then place it at 37°C with 5% CO 2 Cultured in a static incubator for 8 hours (to become a monolayer of cells).

[0084] 2. Take the test solution and dilute to 25 times volume, 250 times volume, 2500 times volume or 25000 times volume with DMEM medium containing 10% (volume ratio) FBS to obtain each dilution.

[0085] 3. Take the cell culture plate that completed step 1, suck and discard the supernatant; add th

[0094] Embodiment 3, the stability of protein solution

[0095] The freshly prepared protein I solution, protein II solution, protein III solution or protein IV solution prepared in Example 1 was placed at room temperature for 3 months, and then used as the test solution (10 parallel treatments were set for each solution).

[0096] 1. Morphological observation

[0097] The protein Ⅰ solution, protein Ⅱ solution and protein Ⅲ solution all became turbid after standing at room temperature for 3 months.

[0098] The protein Ⅳ solution remained clear after 3 months at room temperature.

[0099] 2. Detection of interferon activity

[0100] Method is with embodiment 2. The results are averaged.

[0101] The interferon titer of the protein Ⅰ solution after 3 months at room temperature was 8.2×10 5 U / ml, 48% of freshly prepared.

[0102] The interferon titer of the protein Ⅱ solution after 3 months at room temperature was 3.3×10 5 U / ml, 43% of freshly prepared.

[0103] The in