机构地区: 江苏省农业科学院
出 处: 《农业生物技术学报》 2005年第2期175-178,共4页
摘 要: 参照猪传染性胸膜肺炎放线杆菌(Actinobacilluspleuropneumoniae)血清5型菌株序列Z46774设计1对特异性引物,用PCR方法扩增转铁结合蛋白2(Tbp2)基因,得到1条1624bp的片段,然后克隆到pMD18-T载体中,经测序比较,与参考序列的核苷酸同源性达99.8%,从T-载体中切下目的片段后,定向克隆到pET32a(+)中,转化大肠杆菌(Escherichiacoli)BL21(DE3),经诱导后,SDS-PAGE电泳,Tbp2高效表达,Western-blotting鉴定呈阳性。 To amplify transferrin-binding protein 2 (Tbp2) gene by PCR from Actinobacillus pleuropneumoniae (App), a pair of primers were designed according to sequence Z46774 of A. pleuropneumoniae serotype 5. The amplified DNA fragment 1 624 bp was cloned into pMD18-T and sequenced. The result of sequencing showed that the homology was 99.8% comparing with reference sequence. The target fragment of the positive clones was inserted into pET-32a and transformed in Escherichia coli BL21 (DE3). After induced, the fragment in E.coli BL21 was expressed, the result of Western blotting showed positive.
关 键 词: 胸膜肺炎放线杆菌 铁结合蛋白 传染性 基因克隆 序列分析 电泳 核苷酸同源性 猪 特异性引物 方法 参考序列 载体 定向克隆 大肠杆菌 高效表达 载体 片段 血清 测序