猪博卡病毒(重庆株)NP1部分基因的克隆测序及生物信息学分析

Cloning and Bioinformatics Analysis of Partial NP1 Gene of PBoV Strains from Chongqing

通过设计特异性引物建立基于猪博卡病毒（Porcine Bocavivus,PBoV）NP1基因的特异性PCR方法,从重庆地区猪血液中扩增出PBoV-NP1的部分基因片段,进行基因测序,并与GenBank公布的PBoV、HBoV及PPV基因序列进行生物信息学分析.结果显示：本次试验克隆得到的PBoV重庆株NP1部分基因片段大小为257 bp,与已公布的PBoV-NP1基因的同源性为97.3％～100％;重庆株与PBoV瑞典株、PBoV中国株属同一进化分支,且PBoV重庆株NP1基因与PBoV瑞典株NP1基因亲缘性非常接近;PBoV重庆株NP1基因与GenBank公布的猪PBoV-NP1和1个人HBoV st1聚在博卡病毒一大分支,与PPV分属两大分支,而HBoVst1株又单独形成另一簇.这表明,PBoV重庆毒株与国外毒株的亲缘性非常接近,推测重庆地区猪群中感染的PBoV可能是通过引进或者进口国外动物性产品而传播感染猪群.

Based on the specific PCR of PBoV （Porcine Bocavivus）NP1 gene, we amplified part of the PBoV-NP1 gene from the blood of pigs in chongqing area using specific primer. After sequencing the acquired fragments, we did bioinformatic analysis compared with PBoV, HboV and PPV gene published by the GenBank. The result showed that the length of the PBoV-NP1 genetic frag- ments was 257 bp, and the similarity with the published PboV-NP1 was 97.3%～100%.Phylogenetic tree analysis indicated that the acquired Chongqing PBoV-NP1 fragments in this experiment gathered in the same branch with Sweden PBoV-NP1 strains and China PBoV-NP1 strains, and the Chongqing PBoV-NP1 fragments exhibited a very close evolutionary distance with Sweden PBoV-NP1 strains. Besides, Chongqing PBoV-NP1 strains gathered in the big branch with published pig PBoV-NP1 and one human hHBoV stl. The PPV was in another branch while HBoV stl strain form a cluster by itself. This suggested that Chongqing PBoV-NP1 strains had a very close relationship with Swenden PBoV-NP1 strains, predicting that the infection of PBoV in Chongqing area may caused by im- porting foreign animals or animal products.