马链球菌马亚种IgG结合蛋白的原核表达和免疫原性

邵俊高1 姜慧娇1 常建新1 张宝江1 李善春1 苏艳1

(1.新疆农业大学动物医学学院, 新疆乌鲁木齐 830052)

【摘要】为研究马链球菌马亚种IgG结合蛋白(EAG)免疫原性和保护力,评价其作为马链球菌疫苗抗原的价值。采用PCR法扩增马链球菌马亚种EAG基因,将测序正确的EAG扩增产物与原核表达载体pET-28a(+)连接构建重组质粒,对转化后的大肠杆菌进行诱导表达,用诱导纯化后的重组蛋白作免疫原免疫小鼠,分析重组蛋白免疫小鼠后的抗体水平及对小鼠的免疫保护力。结果表明,诱导后可得到26 kDa的EAG重组蛋白,且该蛋白可与该菌阳性血清发生特异性反应。间接ELISA检测免疫小鼠的抗体效价可达1∶8 100,重组蛋白免疫后对小鼠保护力可达90%。该结果表明,表达的EAG蛋白具有良好的抗原性,可有效提高小鼠的体液免疫水平及免疫保护力。

【关键词】 马链球菌马亚种; EAG蛋白; 表达; 免疫原性分析;

【DOI】

【基金资助】 国家科技支撑项目(No.2012BAD46B01)资助 National Science and Technology Support Program(No.2012BAD46B01)

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This Article

ISSN:1000-3061

CN: 11-1998/Q

Vol 32, No. 05, Pages 577-583

May 2016

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摘要

  • 1 材料与方法
  • 2 结果与分析
  • 3 讨论
  • 参考文献