【摘要】 参考GenBank鸡正呼肠孤病毒(Avian reovirus,ARV)和番鸭呼肠孤病毒(Muscovy duck reovirns,DRV)σ非结构蛋白(σNS)基因序列设计合成1对引物,对番鸭呼肠孤病毒YB(DRV-YB)株σNS基因进行RT-PCR扩增,克隆到pMD18-T载体中,并对克隆产物进行PCR鉴定和测序;DRV-YB株编码σNS的基因全长为1 191bp,其5’和3’端具有典型的禽正呼肠孤病毒的特征,开放阅读框从24～1 127位碱基,编码367个氨基酸残基。DRV-YB株与法国番鸭呼肠孤病毒89026(DRV-89026)株和鸡呼肠孤病毒S1133(ARV-S1133)株σNS基因核苷酸同源性分别为87.3%和76.5%;推导氨基酸同源性分别为94.8%和90.5%。进化树分析表明DRV-YB株σNS与DRV-89026株亲缘关系较近,处在番鸭呼肠孤病毒的分支上。分析发现DRV-YB株S组基因大小和编码蛋白与法国番鸭呼肠孤病毒89026、89330株S组一致,具有法国番鸭呼肠孤病毒89026、89330株S组的特征,而与ARVS1133、176等鸡源呼肠孤病毒差异较大。表明不同禽正呼肠孤病毒株S组基因的大小和编码同一蛋白的等位基因呈现多态性,自然界中禽正呼肠孤病毒不同毒株之间存在基因交换和基因重组现象。更多还原

【Abstract】 The σNS gene of muscovy duck reovirus YB(DRV-YB)was amplified by RT-PCR with oligonucleotide primers,which were designed based on previously reported σNS sequences of avian and muscovy duck reoviruses in the GenBank.The PCR product was cloned into the pMD18-T vector and confirmed by PCR and sequencing analyses.The results show that the σNS gene of DRV-YB contains 1 191 bp,with conserved 5’ and 3’ terminal motifs typical of avian reoviruses.It contains one open reading frame from nt 24 to nt 1 127,which encodes a protein of 367 amino acids.Further nucleotide and amino acid sequence alignments among DRV-YB,DRV-89026,and ARV-S1133 indicate that the σNS nucleotide sequence of DRV-YB is 87.3% and 76.5% homologous to the corresponding genes in DRV-89026 and ARV-S1133,while the deduced amino acid sequence of DRV-YB is 94.8% and 90.5% homologous to DRV-89026 and ARV-S1133,respectively.Phylogenetic analysis shows that the σNS genes of DRV-YB and DRV-89026 are closely related to and fall into the same evolutionary cluster.DRV-YB and DRV-89026 as well as DRV-89330 share the same characteristics of S-class genes,with the same gene length and encoded proteins.However,the genome organization of DRV-YB is dramatically different from those of ARV-S1133 and ARV-176,which are strains from chicken.The results suggest that the S-class genes exhibit polymorphism in the length of gene segments and encoded proteins among different Avian orthoreovirus strains.This polymorphism may reflect exchange and recombination of gene segments among different Avian orthoreovirus strains in nature.更多还原