牦牛病毒性黏膜病病毒P125基因的克隆及序列分析

试验参考GenBank中发表的牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)毒株基因组序列设计了5对引物,利用PCR扩增出预期的3475 bp目的片段;将扩增产物连接至pMD19-T载体,经质粒PCR鉴定及双酶切鉴定获得阳性重组质粒并进行核苷酸序列测定。经BLAST同源性分析表明,牦牛BVDV P125基因与BVDV-Ⅰ型的NADL毒株P125基因的同源性为63.4%。系统分析结果表明,牦牛P125基因与BVDV标准毒株P125基因在遗传进化与亲缘关系上均较远,这可能是因为环境诱变的结果或该毒株具有新的遗传衍化来源。

Cloning and Bioinformatics Analysis of the P125 Gene of Bovine Viral Diarrhea Virus Strain Yak

According to the sequence data of BVDV strain published by GenBank, five set of primers were designed and used to amplify P125 gene of BVDV strain yak by the method of PCR. A specific 3475 bp DNA segment was amplified, which was cloned into pMD19-T vector. The positive recombinant clone was identified by plasmid PCR and restriction enzyme digestion. Compared with BLAST, the P125 gene of BVDV strain yak exhibited the highest homology with strain NADL, but they only shared 63.4%, showing that the P125 gene of BVDV strain yak had great variation. This might be the virus to adapt to yak and the ecological environment of the plateau,or the virus might also have an independent source of genetic.