猪胸膜肺炎放线杆菌16S rDNA基因的克隆及序列分析

Cloning and Sequence Analysis of 16S rDNA Gene of Actinobacillus pleuropneumoniae

为了阐明猪胸膜肺炎放线杆菌16S rDNA基因的遗传变异情况,选择猪胸膜肺炎放线杆菌(Actinobacilluspleuropneumoniae,APP)的16S rDNA基因设计引物,应用聚合酶链反应(PCR)扩增APP的16S rDNA基因的部分序列,将该产物克隆到pMD18-T载体上,重组质粒通过菌落PCR鉴定后,对阳性菌落进行序列测定并进行序列分析。结果表明:本研究得到的长沙(CS)株与血清3、5、7型同源性为99.4%～99.6%,与李氏放线杆菌、脲放线杆菌、马驹放线杆菌、荚膜放线杆菌同源性分别为97.8%、96.3%、97.3%、96.3%。该结果为APP分子流行病学调查、亲缘性关系的研究提供了依据,同时为APP的监控奠定了基础。

This study aimed to clarify the genetic variation situation of 16S rDNA of Actinobacillus pleuropneumoniae （APP）. The 16S rDNA gene of APP was selected to design primers, the partial sequences of 16S rDNA gene of APP were amplified by using polymerase chain reaction （PCR）, and the obtained products were cloned into pMD18-T vector. After the recombinant plasmids were identified by colony PCR, the positive colonies were sequenced and analyzed. The results showed that the homology between obtained Changsha strain （CS） and serum 3, 5, 7 type was 99.4%-99.6%; the homology between CS and A ctinobacillus lignieresii, A ctinobacillus urea, A ctinobacillus equuli, A ctinobacillus capsulatus was 97.8%, 96.3%, 97.3%, 96.3%, respectively. These results provide basis for studies on molecular epidemiology and phylogenetic relationship of APP, as well as laid the foundation for monitoring APP.