马链球菌马亚种新疆分离株XZ1SeM蛋白表达和免疫原性分析

Expression and Analysis on Immunogenicity of SeM Protein of Streptococcus equi subspecies equi XZ1 Strain from Xinjiang

为研究马链球菌马亚种新疆分离株XZ1SeM重组蛋白的免疫生物学功能,评价其作为抗原开展研制马链球菌疫苗的价值。采用PCR方法从马链球菌马亚种扩增其SeM基因,将其克隆至原核表达载体pGEX-4T-2,诱导表达并用纯化的重组蛋白作免疫原免疫小鼠,分析比较重组蛋白免疫后小鼠抗体水平及对小鼠的免疫保护力。结果表明,纯化得到64ku的SeM重组蛋白,且该蛋白可与马链球菌马亚种阳性血清发生特异性反应。间接ELISA检测的抗体效价高达1∶128 000,具有良好的免疫原性,免疫小鼠攻毒结果显示重组蛋白免疫对小鼠有良好的保护效果。

The immunological and biological activity of SeM recombinant protein of Streptococcus equi subspecies equi XZ1 Strain isolated in Xinjiang were studied in order to assess the value of SeM of S.equi for antigen development.The PCR method was applied to amplify SeM.The SeM gene was cloned into pGEX-4T-2prokaryotic expression vector,and induced to express.The purified recombinant protein SeM were used to immunize mice.The antibody level for SeM immunization group and immunological protection of mice were analyzed after immunization.The result showed that there were 64 ku SeM recombinant protein in the purification,and there could be pecullear reaction between the protein and the positive blood serum of S.equi.The antibody titer were 1∶128 000,which had good immunological effects.In addition,this recombinant protein had good protective effects for the mice after immunization.