猪流产衣原体CP/12株omp-1基因的克隆与表达  

Cloning and prokaryotic expression of omp-1 of Chlamydophila abortus CP/12 of swine

作　　者：杨琪[1] 何诚[1] 雷鸣[1] 杨利[1] 朱虹[2] 端青[2] 

机构地区：[1]中国农业大学动物医学院,北京100094 [2]军事医学科学院微生物及流行病学研究所,北京100077

出　　处：《中国农业大学学报》2005年第3期56-59,共4页Journal of China Agricultural University

基　　金：国家自然科学基金资助项目(30370070);北京市自然科学基金资助项目(6052014)

摘　　要：为观察重组MOMP蛋白的免疫活性,揭示omp1基因(编码MOMP蛋白)在猪流产衣原体诊断和防治中的作用,用PCR法扩增猪流产衣原体omp1基因,将特异性扩增片段克隆入pMD18T载体,测序比较分析序列后确认为目的基因,与GenBank中4株流产衣原体omp1基因和氨基酸序列的相似性均达99%,与其他各型衣原体代表株的相似性为71%～88%,氨基酸序列也有较大差异。将omp1基因亚克隆到pET32a表达载体上,再对重组表达载体转化BL21(DE3)表达宿主菌,以IPTG诱导该重组菌,结果显示该重组菌表达了融合蛋白;用猪流产衣原体多克隆抗体对表达的重组MOMP蛋白进行Westernblot试验,结果显示该重组MOMP蛋白具有结合特异性抗体的活性。In order to manifest the function of the omp-1 in preventing and diagnose of swine (strain cp/12), we cloned and expressed the omp-1 in Ecoli. The gene was amplified by PCR. The specific product was cloned into the pMD18-T Vector. After sequence analysis, it was confirmed to be the target gene. Compared with 4 strains of Chlamydophila abortus in GenBank, the homologies of nucleotide sequences of omp-1 and its putative protein sequence were 99%, but 71%-88% similar with other typical strains of Chlamydia psittaci. The gene was subcloned into pET-32a vector. The recombinant vector was transformed into the host bacteria BL21 (DE3). Induced by IPTG, the bacteria with recombinant vector expressed 58ku fusion proteins. Western-blot analysis showed that the protein could react specifically with Chlamydophila abortus polyclonal antibody.

关 键 词：猪流产衣原体 主要外膜蛋白 克隆 表达 

分 类 号：S851.67[农业科学—预防兽医学]