牛支原体广西分离株P48的原核表达及蛋白纯化研究

Study on Prokaryotic Expression and Protein Purification of Mycoplasma Bovis Strain P48 Isolated from Guangxi

为获得牛支原体P48蛋白,以提供免疫学诊断产品及疫苗开发原材料,研究运用PCR方法扩增牛支原体广西分离株的脂蛋白P48的编码基因,构建pET32a-P48重组质粒,导入BL21感受态细胞,在诱导剂异丙基硫代半乳糖苷(IPTG)的诱导作用下获得P48重组蛋白并进行纯化,然后以纯化蛋白免疫小鼠,借助间接ELISA测定血清抗体效价。结果显示,成功构建了P48重组蛋白表达载体,纯化的P48重组蛋白能刺激小鼠产生良好的免疫反应,三免后血清效价达到6.25×10^(5)。

In order to obtain Mycoplasma bovis P48 protein and provide immunological diagnostic products and vaccine development raw materials,the coding gene of lipoprotein P48 of Mycoplasma bovis Guangxi isolate was amplified by PCR,pet32a-P48 recombinant plasmid was constructed,introduced into BL21 competent cells,and P48 recombinant protein was obtained and purified under the induction of inducer isopropyl thiogalactoside(IPTG),Then the mice were immunized with the purified protein,and the serum antibody titer was determined by indirect ELISA.The results showed that the expression vector of P48 recombinant protein was successfully constructed.The purified P48 recombinant protein could stimulate good immune response in mice,and the titer of the serum reached6.25×10^(5) after the third immunization.