摘要
根据GenBank中猪圆环病毒 2型 (PCV 2 )基因序列 ,设计两对特异性引物 ,用PCR方法从接种疑似PMWS仔猪病料的细胞中分段扩增 2个分离毒株全基因组。将扩增片段克隆入 pGEM Teasy载体 ,筛选获得含有相应片段的阳性重组质粒。对质粒中的插入片段进行测序拼接 ,获得 2株均为 176 8bp的全基因组序列。应用DNAstar序列分析软件 ,对所测PCV 2序列与GenBank中的国内外PCV毒株进行同源性比较 ,并绘制系统发生树 ,结果发现 :所测两毒株之间的核苷酸序列同源性极高 ,可达 99 8% ,亲缘关系密切 ;与PCV 2参考毒株的同源性介于95 3%～ 99 7%之间 ,其中与美国的一株PCV 2同源性最高 ,分别为 99 5 %和 99 7% ,亲缘关系很近 ;与国内两分离株同源性分别为 96 4%和 98 8%～ 98 9% ;与PCV 1参考毒株同源性仅为 77 1%～ 77 5 %。
According to the genomic sequence of PCV-2,two sets of PCR primers were designed and two PCV-2 complete genomes were amplified by PCR from RK and Vero cells inoculated with PMWS samples.The PCR products were cloned into pGEM-T easy vector and the recombinant plasmids were obtained.The sequences of the cloned PCR products were analysed and the complete genomes of two PCV-2 isolates were determined,both of them are 1768bp in length.The two sequences were compared with the other PCV isolates in GenBank,the results indicated that the two PCV-2 isolates sequenced are closely related to each other,displaying 99.8% nucleotide sequence identity,having 95.3%～99.7% sequence identity with the other PCV-2 and shared the most high sequence identity with an USA isolate,99.5% and 99.7% respectively.Moreover the two isolates have 96.4% and 98.8%～98.9% sequence identity with two other Chinese isolates and there exists only 77.1%～77.5% nucleotide identity with PCV-1 isolates.
出处
《病毒学报》
CAS
CSCD
北大核心
2003年第1期42-46,共5页
Chinese Journal of Virology
基金
国家 8 6 3高技术发展计划资助项目 (批准号 :2 0 0 1AA2 490 12 )
上海市科技兴农攻关项目 (批准号 :农科攻字( 2 0 0 1)第 3 -5号 )
