| 胸膜肺炎放线杆菌ApxIA N端基因的克隆与原核表达 |
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| 引用本文： | 吴笳笛,苏禹刚,费东亮.胸膜肺炎放线杆菌ApxIA N端基因的克隆与原核表达[J].安徽农业科学,2009,37(6):2419-2421. |
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| 作者姓名： | 吴笳笛 苏禹刚 费东亮 |
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| 作者单位： | 辽宁医学院畜牧兽医学院,辽宁锦州,121000;辽宁医学院畜牧兽医学院,辽宁锦州,121000;辽宁医学院畜牧兽医学院,辽宁锦州,121000 |
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| 摘 要： | 目的]为猪接触传染性胸膜肺炎的诊断和基因工程疫苗的研制提供理论依据。方法]以胸膜肺炎放线杆菌（APP）血清10型菌株为材料,采用PCR技术对其ApxIA的N端基因进行扩增,将扩增产物转入克隆载体和表达载体中进行克隆和表达,用重组表达质粒转化大肠杆菌BL21（DE3）,利用免疫印迹分析鉴定ApxIAN端基因表达产物的免疫活性。结果]ApxIA N端基因PCR扩增产物与标准10型ApxIAgene（D16582）的同源性为99.7%。NcoI和HindⅢ双酶切鉴定结果表明,目的基因已成功转入原核表达载体pET-32a中。含重组质粒的大肠杆菌BL21经IPTG诱导后表达了相对分子量为779 kDa的目的蛋白,该蛋白分子量与标准ApxIA蛋白（Marker）相同。结论]SDS-PAGE电泳和免疫印迹检测结果表明,ApxIAN端基因在表达载体中得到高效表达,且表达产物具有免疫活性。
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| 关 键 词： | 胸膜肺炎放线杆菌 毒素IA 克隆 检测 |
Cloning and Prokaryotic Expression of ApxIA Gene of Actinobacillus pleuropneumoniae |
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| WU Jia-di et al.Cloning and Prokaryotic Expression of ApxIA Gene of Actinobacillus pleuropneumoniae[J].Journal of Anhui Agricultural Sciences,2009,37(6):2419-2421. |
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| Authors: | WU Jia-di |
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| Institution: | WU Jia-di et al(Animal Husb,ry , Veterinary Medicine College,Liaoning Medical University,Jinzhou,Liaoning 121000) |
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| Abstract: | Objective]The study was to provide the theoretical basis for diagnosis on pig contact infectious pleuropneumoniae and development of genetic engineering vaccine.Method] With 10 type strain of Actinobacillus pleuropneumoniae(APP) serum as the material,PCR technique was taken to amplify N-end gene of ApxIA,and the amplified products was transferred into cloning and expression vector to clone and express And the recombinant expression plasmid was taken to transform Escherichia coli BL21(DE3),the immune activ... |
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| Keywords: | Actinobacillus pleuropneumoniae Toxin IA Cloning Detection |
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