| Theileriosis of Bovine is caused by Theileria orientalis which evoked the livestock high fever, anemia, jaundice, and infected with some other virus and parasites that can induced the livestock deceace on occasions. There wasn’t an effective prevention and cure method for Theileriosis nowdays.The p23gene(EU573168.1) and p33gene(DQ078264.1) of MPSP (Major piroplasm surface protein) sequence were cloned to pMD18-T Simple Vector, the prokaryotic expression vector pETpET30a-p23and pETpET30a-p33were constructed. The size and reactogenicity of the expressed proteins were analyzed by Western blot. The optimum concentration of IPTG, the optimum culture medium and the optimum time for induced were screened. The levels of the specific antibody and the cytokines(IL-2, IFN-γ and IL-4) in the serum of the each group immunized BALB/c mice and Bovine. The atlas of T-LympHocyte Subsets were evaluated in different groups of immunized BALB/c mice.The results showed that the prokaryotic expression vector pETpET30a-p23and pETpET30a-p33were constructed, the expressed fusion protein was24kDa,31kDa, respectively. Both of the fusion protein have a reactogenicity in Western blot test. The optimum concentration of IPTG and the optimum induced time was0.1mM with6h in common, The optimum culture medium for recombinant plasmid pET30a-p23and pET30a-p33was LB and SOC respectively.100μg with each BALB/c mice was the maximum security dosage. The Co-immunization group has a significant difference (P<0.05) with the single immunization group of BALB/c mice and Bovine in levels of the specific antibody and the cytokines(IL-2, IFN-γ and IL-4), Same as the content analysis of T-LympHocyte Subsets. The Co-immunization method with recombinant protein vaccine pET30a-p23and pET30a-p33induced a better immune effect than which pET30a-p23only or pET30a-p33only in immuned BALB/c mice and cattle group. And this provide a new sight for the prevention of Bovine Theileriosis. |
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