猪流产衣原体CP/12株omp-1基因的克隆与表达

为观察重组MOMP蛋白的免疫活性，揭示omp-1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omp-1基因，将特异性扩增片段克隆入pMD18-T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omp-1基因和氮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omp-1基因亚克隆到pET-32a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Western blot试验，结果显示该重组MOMP蛋白具有结合特异性抗体的活性。

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

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.