| 副猪嗜血杆菌外膜蛋白TbpB基因的克隆和表达 |
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| 引用本文： | 叶飞,张培君,田德雨,孙慧玲,王宏俊,龚玉梅,贺云霞.副猪嗜血杆菌外膜蛋白TbpB基因的克隆和表达[J].黑龙江畜牧兽医,2011(15). |
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| 作者姓名： | 叶飞 张培君 田德雨 孙慧玲 王宏俊 龚玉梅 贺云霞 |
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| 作者单位： | 北京市农林科学学院畜牧兽医研究所;首都师范大学生命科学学院;中国农业大学动物医学院; |
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| 基金项目： | 国家自然科学基金项目(31001072); “863”国家高技术研究发展计划项目(2006AA10A206); 北京市农林科学学院青年基金项目(QNJJ201012),北京市农林科学学院一般项目(2010A008) |
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| 摘 要： | 为了克隆和表达副猪嗜血杆菌(Haemophilus parasuis,HPS)外膜蛋白TbpB基因,试验采用基因工程的方法对已发布的HPS外膜蛋白P5序列进行相关序列分析,设计并合成引物,以HPS血清5型基因组为模板,通过PCR扩增HPS外膜蛋白P5编码基因,获得长为1 116 bp的预期基因片段。该基因编码372个氨基酸的蛋白质,预测分子质量约为41 ku,将该基因片段定向克隆至表达载体pGE X-6p-1中。结果表明:诱导表达后分子质量约为67 ku,与副猪嗜血杆菌阳性血清发生了特异性反应。说明该蛋白与抗体发生了反应,有一定免疫原性。
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| 关 键 词： | 副猪嗜血杆菌 转铁结合蛋白B(TbpB) 克隆 表达 |
Cloning and expression TbpB gene for outer membrane protein of Haemophilus parasuis |
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| YE Fei,ZHANG Pei-jun,TIAN De-yu,SUN Hui-ling,WANG Hong-jun,GONG Yu-mei,HE Yun-xia.Cloning and expression TbpB gene for outer membrane protein of Haemophilus parasuis[J].Heilongjiang Animal Science And veterinary Medicine,2011(15). |
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| Authors: | YE Fei ZHANG Pei-jun TIAN De-yu SUN Hui-ling WANG Hong-jun GONG Yu-mei HE Yun-xia |
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| Institution: | YE Fei1,2,ZHANG Pei-jun1,TIAN De-yu3,SUN Hui-ling1,WANG Hong-jun1,GONG Yu-mei1,HE Yun-xia1(1.Institute of Animal Husbandry and Veterinary Medicine,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100097,China,2.College of Life Sciences,Capital Normal University,Beijing 100048,3.College of Veterinary Medicine,China Agriculture University,Beijing 100094,China) |
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| Abstract: | To study the immunogenicity of the proteins related with HPS,A pair of specific primers were designed and synthesized on the basis of the published genome as a template.The TbpB gene of HPS was amplified by PCR,and then was cloned into the expression vector pGEX-6p-1.The fusion protein was successfully expressed,analyzed the immunogenicity of TbpB protein by the Western-blot.The results showed that the TbpB protein was obtained and its molecular weight was about 54 ku.The Western-blot results showed that it... |
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| Keywords: | Haemophilus parasuis(HPS) TbpB cloning expression |
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