禽呼肠孤病毒μNS非结构蛋白基因的克隆及序列分析

Cloning and sequence analysis of the μNS gene of Avian reovirus

根据GenBank中禽呼肠孤病毒(ARV)M3基因序列,设计并合成了一对跨越μNS非结构蛋白基因完整开放阅读框(ORF)的特异性引物,对ARV的10个毒株进行RT-PCR扩增、克隆及序列测定。结果表明,10个毒株μNS蛋白基因ORF的核苷酸序列全长均为1908 bp,编码635个氨基酸。这10个毒株之间的核苷酸及推导的氨基酸同源性分别都在98%以上。将它们与哺乳动物呼肠孤病毒(MRV)、番鸭呼肠孤病毒(DRV)等进行核苷酸及推导的氨基酸序列比较,并进行遗传系统树分析,结果表明ARV与MRV有较大的差异,与DRV差异较小。

A pair of specific primers were designed and synthesized according to M3 gene sequence of Avian reovirus （ARV） from Gen-Bank for the whole Open Reading Frame （ORF） of μNS nonstructural protein gene. The μNS gene of ten ARV strains were amplified, cloned and sequenced. The whole ORF of μNS protein gene were 1 908 bp in length and predicted to encode a μNS nonstructural protein of 635 amino acids. The percentages of nucleotide and amino acid sequence identities among ten ARV strains were all above 98%. Resuts of nucleotide and predicted amino acid sequences and phylogenetic analysis between ARV and other orthoreoviruses （MRV and DRV） showed that there was a signifcunt high divergence between ARV and MRV, but smaller between ARV and DRV.