猪传染性胸膜 炎放线杆菌转铁结合蛋白基因克隆序列分析及表达.pdf

业生物技术学报 Journal of Agricultural Biotechnology 2005,13 (2): 175~ 178 ·研究论文· 猪传染性胸膜肺炎放线杆菌转铁结合蛋白2 基因克隆、 序列分析及表达* 芳 何孔旺** (江苏省农业科学院兽医研究所, 农业部畜禽疫病诊断重点 放实验室,南京2 10014) 摘要：参照猪传染性胸膜肺炎放线杆菌( )血清5 型菌株序列Z46774 设计 1 对特异性引物， 得到 ，与参考序 PCR 方法扩增转铁结合蛋白2( )基因， 1 条 1 624 bp 的片段，然后克隆到pMD18-T 载体中，经测序比较 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( )BL21 ， ， ， 。 (DE3) ，经诱导后 SDS电泳 高效表达 Western-blotting 鉴定呈阳性 关键词：胸膜肺炎放线杆菌; 转铁结合蛋白2; 克隆; 序列分析; 表达 Cloning, Sequencing and Expression of the Transferrin-binding Protein 2 Gene from WANG Fang HE Kong-Wang** ( ) To amplify transferrin-binding protein 2 ( ) gene by PCR from (App), a pair of primers were designed according to sequence Z46774 of serotype 5. The amplified DNA fragment 1 624 bp was cloned into pMD 18-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 BL2 1 (DE3). Af- ter induced, the fragment in BL2 1 was expressed, the result of Western blotting showed positive. ; transferrin-binding protein 2; cloning; sequencing; expression 随着养猪规模化程度的不断提高，猪传染性胸 泛研究。与