A recombinant bacillus calmette-guerin vaccine used for preventing toxoplasmosis in pigs and a preparing method thereof
A technology for recombinant BCG and toxoplasmosis, which is applied in the field of vaccines, can solve problems such as unsatisfactory immune effects of vaccines, and achieve the effects of good immune protection, prevention of toxoplasmosis, and stability and safety.
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Embodiment 1
[0032] 1. Preparation and evaluation of recombinant BCG vaccine for porcine toxoplasmosis prevention with shuttle expression vector:
[0033] (1) Preparation steps of pMV261-IL2-Rho recombinant BCG vaccine for preventing porcine toxoplasmosis:
[0034] 1. Primer design
[0035] According to the DNA sequence of porcine interleukin 2 (IL-2) gene, the DNA sequence of Toxoplasma gondii Rhomboid gene and the vector map of shuttle vector pMV261, 2 pairs of primers were designed and restriction sites were introduced.
[0036] Porcine interleukin 2 (IL-2) upstream primer IL-F: 5'CG GAATTC ATGTATAAGATGCAGCTC 3', where the restriction site at the 5' end is Eco RI.
[0037] Downstream primer IL-R: 5'CGC GGATCC AGTCAGTGTTGAGTAGATGCTTTG3', wherein the restriction site at the 5' end is Bam Hi.
[0038] The upstream primer of Toxoplasma gondii Rhomboid gene is R-F: 5'CGC GGATCC ATGCCTATTCGCTTTCTCG 3', the restriction site at the 5' end is Bam Hi.
[0039] Downstream primer R-R:...
Embodiment 2
[0074] 2. The preparation and evaluation effect of recombinant BCG vaccine integrated with expression vector for porcine toxoplasmosis prevention:
[0075] (1) Preparation steps of pMV361-IL2-Rho recombinant BCG for preventing porcine toxoplasmosis:
[0076] 1. Primer design
[0077] According to the DNA sequence of porcine interleukin 2 (IL-2) gene, the DNA sequence of Toxoplasma gondii Rhomboid gene and the vector map of shuttle vector pMV361, 2 pairs of primers were designed and restriction sites were introduced.
[0078] Porcine interleukin 2 (IL-2) upstream primer IL-F: 5'CG GAATTC ATGTATAAGATGCAGCTC 3', where the restriction site at the 5' end is Eco RI.
[0079] Downstream primer IL-R: 5'CGC GGATCC AGTCAGTGTTGAGTAGATGCTTTG3', wherein the restriction site at the 5' end is Bam Hi.
[0080] The upstream primer of Toxoplasma gondii Rhomboid gene is R-F: 5'CGC GGATCC ATGCCTATTCGCTTTCTCG 3', the restriction site at the 5' end is Bam Hi.
[0081] Downstream primer ...
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