| 副猪嗜血杆菌血清5型TbpA蛋白的表达及其免疫原性分析 |
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| 引用本文： | 李建军,胡明明,朱晓凯,李艳玲,张艳禾,谢芳,王春来.副猪嗜血杆菌血清5型TbpA蛋白的表达及其免疫原性分析[J].中国预防兽医学报,2012,34(3):238-240. |
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| 作者姓名： | 李建军 胡明明 朱晓凯 李艳玲 张艳禾 谢芳 王春来 |
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| 作者单位： | 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/细菌病研究室,黑龙江哈尔滨,150001 |
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| 基金项目： | 兽医生物技术国家重点实验室自主研究课题副猪嗜血杆菌灭活苗的研制(SKLVBP201015) |
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| 摘 要： | 为原核表达副猪嗜血杆菌血清5型TbpA蛋白,本研究利用特异性引物扩增tbpA基因,并将其克隆到pMD 18-T载体中进行序列测定.再将其亚克隆于pET-28a中构建重组表达质粒pET-tbpA.将其转化受体菌E.coli BL21 (DE3),经IPTG诱导后,SDS-PAGE电泳和western blot分析表明,表达的重组蛋白约106 ku,并以包涵体形式存在.表达的重组蛋白经纯化后,免疫6周龄昆明小鼠制备免疫血清,ELISA分析表明制备的血清抗体效价在1∶3 200以上,表明TbpA具有良好的免疫原性.
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| 关 键 词： | 副猪嗜血杆菌 tbpA基因 原核表达 |
Expression of tbpA gene from Haemophilus parasuis serotype 5 and identification of the recombinant protein |
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| LI Jian-jun
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HU Ming-ming
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ZHU Xiao-kai
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LI Yan-ling
,
ZHANG Yan-he
,
XIE Fang
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WANG Chun-lai.Expression of tbpA gene from Haemophilus parasuis serotype 5 and identification of the recombinant protein[J].Chinese Journal of Preventive Veterinary Medicine,2012,34(3):238-240. |
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| Authors: | LI Jian-jun HU Ming-ming ZHU Xiao-kai LI Yan-ling ZHANG Yan-he XIE Fang WANG Chun-lai |
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| Institution: | (State Key Laboratory of Veterinary Biotechnology,Division of Animal Bacteriosis,Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences,Harbin 150001,China) |
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| Abstract: | To express Haemophilus parasuis serotype 5 TbpA protein,the tbpA gene was amplified by PCR and cloned into vector pMD-18T for sequencing.The tbpA gene was subcloned into vector pET-28a.The resultant recombinant plasmid pET-tbpA was transformed into BL21(DE3) and induced with IPTG.The results of SDS-PAGE and western blot indicated that the expressed TbpA protein was about 106 ku and mainly existed in form of inclusion bodies.The anti-tbpA serum was prepared in Kunming mice immunized with purified TbpA with the titer of 1∶3,200,which showed that the express product of TbpA had strong immunogenic reaction. |
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| Keywords: | Haemophilus parasuis tbpA gene prokaryotic expression |
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