鹅细小病毒VP1-VP3非重叠区基因的克隆与序列分析

Cloning and Sequencing of Non-repeated VP1 and VP3 Gene of Goose Parvovirus

通过克隆鹅细小病毒(Goose parvovirus,GPV)分离株VP1-VP3非重叠区基因,并对其进行序列分析,为鹅细小病毒感染与疫苗免疫的鉴别诊断奠定理论基础。进行鹅胚病毒增殖,收集尿囊液,提取基因组,参考发表的B株序列,设计合成一对引物,经PCR扩增,克隆VP1-VP3基因,筛选阳性克隆,对其进行序列测定及同源性分析。结果表明,克隆的基因片段为901 bp,VP1-VP3基因共594 bp,编码198个氨基酸;弱毒株之间亲缘关系很近,核苷酸同源性99.5%～100%, 氨基酸同源性98.5%～100%; 强毒株与B株亲缘关系较近, 核苷酸同源性96.6%,氨基酸同源性97.5%;弱毒株与强毒株亲缘关系较远,核苷酸同源性92%～93%,氨基酸同源性96%～97.5%;弱毒株与B株亲缘关系最远,核苷酸同源性92.5%～93%,氨基酸同源性93.9%～95.5%。说明强毒株与弱毒株之间核苷酸序列存在差异。

A pair of specific primers was designed according to the gene sequence of goose parvovirus（GPV） B strain and used to amplify VP1-VP3 gene by PCR,sequence and analyze for homology.The result of sequencing showed that the product of PCR of VP1-VP3 gene from GPV B strain was consisted of 594 bases,encode 198 amino acids.The homology of nucleotides of four isolated low virulent strains was 99.5% to 100%;the homologies of nucleotides of standard B strains were 96.6% to that of isolated virulent strain,92.5% to 93% to that of isolated low virulent strains.The result showed that a certain variation was observed in the nucleotide and amino acid sequences of low virulent strains and virulent strains of GPV in Heilongjiang province.