绵羊肺炎支原体Y98标准株P30外膜蛋白与IFN-γ的融合表达及其免疫原性

The expression of P30-IFN-γ fusion protein of mycoplasma ovipneumonia(Y-98)and its immunogenicity

以绵羊肺炎支原体(Mycoplasma oumvipneonia,MO)标准株Y98的P30外膜蛋白为抗原,同时以细胞因子IFN-γ为佐剂,将两者串联融合表达。小鼠免疫保护试验结果显示,P30重组蛋白的免疫保护率为60%,IFN-γ重组蛋白的免疫保护率为20%,P30-IFN-γ重组蛋白的免疫保护率达到80%。间接ELISA试验证实,经100μL与200μL P30-IFN-γ重组蛋白免疫小鼠的血清中,P30抗体效价分别为1∶32 000与1∶64 000。以上结果证实,所制备的重组蛋白对小鼠实验个体基本安全,且P30-IFN-γ重组蛋白中的细胞因子IFN-γ组分可以调节机体免疫机能,增强P30蛋白的免疫保护效果。

In this study,the P30 outer membrane protein of mycoplasma ovipneumoniae Y98 as an tigen and IFN ,as vaccine adjuvant,were bound together. The recombination gene was construc ted to fusion expression. After inducing and purifying, we obtained the soluble proteins of gene P30, IFN7 and P30IFN7. The results of immune protect experiment in mice showed that the im mune protective rates of P30 and IFN7 proteins were respectively 600/00 and 20%,while the im mune protective rates of P30IFN7 fusion proteins reached 80 %. Indirect EI.ISA demonstrated that the antibody titer of P30 protein was 1 ： 32 000 in the mice serum from mice injected with 100 /1I. P30IFN7 fusion proteins;while in another mice serum from mice injected with 200 ,uI. P30 IFN7 fusion proteins,the antibody titer was 1 ： 64 000. In conclusion,all the proteins described a bove were safe and effective;moreover, IFN7 could regulate the immune function and enhance the efficiency of recombinant proteins.