番鸭呼肠孤病毒M(M1～M3)基因序列克隆及特性分析

摘要

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2 283 bp,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13～2 211 bp,编码μA蛋白,含732个氨基酸.M2基因序列全长2 155 bp,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28～2 058 bp,编码外壳μB蛋白.M3基因序列全长1 997 bp,而ARVM3基因全长1 996 bp.M3基因仅有1个编码框25～1 683 bp,编码μNS蛋白.MDRV MW9710株M3基因核苷酸序列与MDRV89330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. Ml 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. Ml 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 S14 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. OKd. 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 Ml, M2, M3 genome segment respectively.