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| 1 | 猪传染性胸膜肺炎放线杆菌apxIA基因在大肠杆菌中的融合表达与纯化显示文摘选取猪传染性胸膜肺炎放线杆菌(APP)apxIA基因序列中的抗原决定簇集中的区域,采用PCR方法从APP血清1型参考株259的基因组DNA中,扩增apxIA基因中约954bp的片段,连接到pMD-18T载体,经测序正确后,以EcoRI和NotI双酶切,亚克隆到原核表达载体pGEX-4T-1中,转化大肠杆菌DH5α,经0.4mmol/LIPTG诱导表达,产物通过尿素变性复性,并以Glutathione Sepharose 4B亲和层析的方法对目的蛋白进一步纯化。SDS-PAGE分析结果显示,目的基因在大肠杆菌DH5α中以包涵体形式高效表达,经薄层凝胶扫描分析占菌体总蛋白的32%,纯化后的GST融合蛋白纯度达到95%,为亚单位疫苗和诊断抗原的研究奠定了基础。 | 黄琦 谢芝勋 庞耀珊 刘加波 邓显文 谢志勤 谢丽基 | 2008 | 微生物学通报2008，35，7： | 3 |
| 2 | 利用菌落多重PCR对传染性胸膜肺炎放线杆菌进行血清分型显示文摘参照文献报道的传染性胸膜肺炎放线杆菌的特异基因合成5对特异引物,建立传染性胸膜肺炎放线杆菌血清型分型的菌落多重PCR方法,结果为10株传染性胸膜肺炎放线杆菌血清型参考菌株均扩增出了相应的预期片段,而支气管败血波氏杆菌、多杀性巴氏杆菌、大肠埃希菌的扩增均为阴性。利用此多重PCR方法对41株传染性胸膜肺炎放线杆菌分离菌株进行血清型分型,结果所有菌株均扩增出了相应的特异片段,其中6株为1型,5株为7型,1株为5型,29株为9型。 | 赵耘 杜昕波 李伟杰 陈敏 康凯 | 2010 | 动物医学进展2010，31，1： | 1 |
| 3 | 当前传染性胸膜肺炎的诊断与防控体会显示文摘传染性胸膜肺炎(Actinobacillus pleuropneumo-niae,APP)是由胸膜肺炎放线杆菌感染所致的一种纤维素性、出血性、坏死性肺炎。Shope最早于1964年提出本病相关的报告之后,本病例一直广泛分布于全世界,对全球养猪业造成了巨大经济损失[1],是猪只重要的呼吸道疾病,所有年龄阶段均可感染,但以肥育猪较常见。美国很多猪群已基本净化了本病,但APP目前在我国比较普遍,临床症状与其它呼吸道疾病相似,在临床上易造成误诊,故笔者将自己对APP的诊断与防控措施的体会作如下总结,以期与同行分享。 | 曲向阳 邢军 吴俊 李喜焕 张聪洲 | 2012 | 养猪2012，，4： | 1 |
| 4 | Cloning and Expression of Actinobacillus pleuropneumoniae Gene Coding for TbpA and Development of an Indirect TbpA-ELISA显示文摘This study presents the cloning and expression of gene encoding transferrin-binding protein A from Actinobacillus pleuropneumoniae in Escherichia coli expression system and the development of an indirect TbpA-ELISA. The gene coding TbpA was amplified from the A. pleuropneumoniae serotype 2 genome using polymerase chain reaction and cloned to pET-28b expression vector under the control of strong, inducible T7 promoter. The recombinant plasmid was expressed in E. coli BL21 (DE3). The expressed fusion protein was analyzed using SDS-PAGE and Western blotting. The diagnostic potential of recombinant TbpA (rTbpA) was evaluated through an antibody-detection indirect ELISA based on the purified rTbpA. The TbpA antibodies were detectable in mice on day 7 after vaccination with purified rTbpA protein or infection with A. pleuropneumoniae serotype 10 with the TbpA-based ELISA. In addition, the TbpA-ELISA was able to detect 12 serotyping rabbit antisera postinoculation (PI) with A. pleuropneumoniae 12 serotypes experimentally. The comparable result was obtained by detecting the 117 clinical serum samples using, respectively, the TbpA-ELISA and indirect hemagglutination test (IHA) based on multiplex antigen. The result indicates that the TbpA-ELISA was the more sensitive method compared with the Mix-IHA method because of its consistent presence in A. pleuropneumoniae serotypes. In conclusion, the conserved TbpA of A. pleuropneumoniae can be used for the development of a cross-serotype diagnostic method for the detection of antibodies against A. pleuropneumoniae. | LIANG Wang-wang HE Qi-gai CHEN Huan-chun XU Di-ping WU Rui ZHANG Rong-rong | 2008 | Agricultural Sciences in China2008，7，10： | 0 |
| 5 | 猪传染性胸膜肺炎放线杆菌毒素Ⅲ A基因的克隆及其在巴斯德毕赤酵母中的分泌表达显示文摘应用PCR方法扩增猪传染性胸膜肺炎放线杆菌(Actinabacillus pleuropneumoniae,App)编码毒素A结构蛋白的完整基因,连接到载体pMD18-T中,经Not和Snab双酶切后连接到用同样酶切的表达载体pPIC9k上,转化GS115酵母感受态细胞,经甲醇诱导表达了分泌到胞外的Apx A蛋白。SDS-PAGE分析显示,表达的蛋白约为110 000,与预期相符。Dot-blot和ELISA检测结果表明,表达蛋白具有良好的抗原性和特异性。为进一步研究Apx A蛋白的结构和功能,研制App重组诊断抗原或亚单位疫苗奠定了基础。 | 汪招雄 何启盖 刘军发 刘正飞 陈焕春 | 2007 | 中国兽医学报2007，27，6： | 0 |
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