番鸭源小鹅瘟病毒PT株VP基因的克隆与序列分析

为了获得番鸭(Cairina moschata)源小鹅瘟病毒(Goose parvovirus,GPV)结构蛋白VP基因的相关信息,根据国内外已发表鹅源GPV与番鸭细小病毒(Muscovy duck parvovirus,MDPV)全基因序列,应用DNAStar分子生物学软件设计一对引物,应用高保真PCR技术扩增番鸭源小鹅瘟病毒PT株(GPV PT株)VP全基因序列.将扩增得到的VP全基因克隆到pMD 18-T载体上,获得的重组质粒经PCR鉴定后进行序列测定.结果表明,PT株VP全基因大小为2 199 bp,编码732个氨基酸(GenBank登录号:JF926695),与番鸭源小鹅瘟病毒GPV DY株核苷酸及其推导氨基酸同源性分别为98.8％和98.8％,高于鹅源GPV与MDPV参考毒株.在国内外首次发现番鸭源GPV VP基因VP1独特区具有MDPV核苷酸序列特征,而VP2基因具有鹅源GPV核苷酸序列特征.本研究从番鸭源GPV PT株成功克隆到结构蛋白全基因序列,为深入研究番鸭源GPV的起源及水禽细小病毒遗传衍化提供参考.

Cloning and Sequence Analysis of VP Gene of Goose parvovirus PT Strain Isolated from Muscovy Duck Origin

To characterize the structural protein VP gene of muscovy duck(Cairina moschata) origin Goose parvovirus(GPV) PT strain,a pair of primers were designed by DNAStar software based on the published sequences of goose original GPV and Muscovy duck parvovirus(MDPV) in GenBank.High fidelity PCR was performed,and the obtained PCR products were cloned into the pMD18-T vector,sequenced and analyzed.Sequencing analysis demonstrated that the VP gene of GPV PT strain included 2 199 bp encoding 732 amino acids(GenBank number:JF926695).The VP gene of GPV PT strain showed a combined feature of GPV and MDPV.It shared a nucleotide identity of 98.8% and an amino acid identity of 98.8% with muscovy duck origin GPV DY strain,which were much higher than the identities with that of goose origin GPV and MDPV.The VP1 gene unique part was similar to that of MDPV and the VP2 gene had the characteristic of goose origin GPV,which have not been reported previously.The success of this study has provided clues for the origin of muscovy duck origin GPV,and even make it possible to study the evolution and phylogenesis of Waterflow parvovirus.