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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;

Prokaryotic expression and immunogenicity of IgG-binding protein of Streptococcus equi subspecies equi

Jungao Shao1 Huijiao Jiang1 Jianxin Chang1 Baojiang Zhang1 Shanchun Li1 Yan Su1

(1.College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi, Xinjiang, China 830052)

【Abstract】To analyze the immunogenicity and protective ability of recombinant IgG-binding protein (EAG) of Streptococcus equi subspecies equi and to evaluate its value when used as equine vaccine antigen, EAG gene was amplified by PCR and inserted into pET-28a vector. The EAG recombinant proteins were expressed and purified to immune mice. The serum antibody and challenge protection were tested. The purified recombinant protein of EAG was 26 kDa, and the protein reacted specifically with positive serum of Streptococcus equi subspecies equi. The mice antibody level for EAG immunization group was 1:8 100. The immunological protection result showed that the protection rate of the EAG recombinant protein was 90%. The results suggested that the EAG protein has good immunogenicity and immunological protection, and it can effectively increase the humoral immune response and immunological protection of mice.

【Keywords】 Streptococcus equi subspecies equi; EAG protein; expression; immunogenicity analysis;

【DOI】

【Funds】 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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Abstract

  • 1 Materials and methods
  • 2. Results and Analyses
  • 3 Discussion
  • References