番鸭呼肠孤病毒M(M1～M3)基因序列克隆及特性分析

Cloning and Characteristic Analysis of M Class Genome Segments in Muscovy Duck Reovirus MW9710

参考Genbank番鸭呼肠孤病毒(muscovy duck reovirus,MDRV)中片段基因序列设计合成引物,对MDRV MW9710株中片段M基因RT-PCR扩增后,进行测序和特性分析。结果显示MW9710株M1基因全长2 283bp,与MDRV S14株同源性为99.9%,与ARV同源性小于74%。M1基因仅有1个编码框13～2 211bp,编码μA蛋白,含732个氨基酸。M2基因序列全长2 155bp,与MDRV S14株同源性为99.9%,与ARV同源性小于69%。M2基因仅有1个编码框28～2 058bp,编码外壳μB蛋白。M3基因序列全长1 997bp,而ARVM3基因全长1 996bp。M3基因仅有1个编码框25～1 683bp,编码μNS蛋白。MDRV MW9710株M3基因核苷酸序列与MDRV 89330株的同源性为87.2%,与ARV同源性小于74%。MDRV MW9710株5′末端序列不保守:M1为5′-ACUUUUU,M2为5′-UCUUUUU,M3为5′-GCUUUUU,MDRV MW9710株3′末端序列UCAUC-3′保守。

The primers were designed and synthesized according to the reported muscovy duck reovirus （MDRV） M gene in Genbank. M gene in muscovy duck reovirus MW9710 strain were amplified by RT-PCR and went on sequencing and characteristic analysis. M1 genome segment was 2283bp in length, the nucleotide sequence had 99.9% homology with MDRV S14 strain and no more than 74.0% homology with ARV. M1 gene contained only one ORF （13bp-2211bp） that was predicted to encode μA protein of 732 amino acid. M2 genome segment was 2155bp in length, the nucleotide sequence had 99.9% homology with MDRV 814 strain and no more than 69.0% homology with ARV. M2 gene contained only one ORF （28- 2058bp） that was predicted to encode μB protein of 676 amino acid. M3 genome segment was 1997bp in length and the sequence had 95.3% homology compared to MDRV 89330, but at most 74% homology with ARV. M3 gene in muscovy duck reovirus contained an ORF （25- 1683bp） that encoded a protein of 552 amino acids with a molecular mass of 60. 0Kd. but ARV lacked a nucleotide C in 1169bp, that made the ORF （25 - 1932bp） encode 635 amino acids. The 3＇ terminal conserved motif was UCAUC-3＇, but 5＇terminal conserved motif was 5＇ACUUUUU, 5＇UCUUUUU, 5＇GCUUUUU in M1, M2, M3 genome segment respectively.