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
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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ARV M3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。Abstract: 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 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 60.0Kd. 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.