Cloning And Sequence Analysis Of The M Genes Of Avian Reovirus

According to the M genes (M1,M2and M3)sequence of ARV strains in GenBank, four pairs of primers were designed and synthesized. The RNA of Avian reovirus strains C-98 and T-98 isolated in Inner Mongolia and Tianjin were extracted, respectively. The M genes of revirus were amplificated with specific primers by RT-PCR,then cloned into PMD19-T vectors and certified by PCR, restriction endonucleases analysis and sequenced. The M1 gene segments were 2283 base pairs long, encoding a proteinμA consisted of 732 amino acids. M2 or M3 genome segments were 2158 or 1996 base pairs long, respectively, encoding a proteinμB orμNS consisted of 676 and 635 amino acids, respectively. The genome segments have the 5' GCTTTT,TCATC3' terminal motif which was common to other known avian reovirus genome segments. Non coding regions of both 5' and 3' terminal of the M1 genome segments consist of 12 and 72 nucleotides,respectively, those of each M2 genome segments consist of 29 and 98 nucleotides, respectively, and those of each M3 genome segments were 24 and 64 nucleotides, respectively.Alignment of the homologous ARV C-98,ARV T-98 and other ARVs M1 nucleotide sequences revealed relatively high overall nucleotide identity.The homology of ARV C-98,ARV T-98 and ARV S1133,ARV 1733,ARV 2408 M1 gene were 99.5%～99.7%, The homology of ARV C-98,ARV T-98 and ARV 176 M1 gene were 98.1%～98.2%, The homology of ARV C-98,ARV T-98 and ARV OS161,ARV138,ARV1017-1,ARV 918, ARV 916SI M1 gene were 88.0%～92.2%,The homology of ARV C-98,ARV T-98 and DRV S14 M1 gene were 69.5%～69.7%, The low homology of ARV C-98,ARV T-98 and MRV T1L,MRV T3D M1 gene were 28%, respectively.The homology of ARV C-98,ARV T-98 and ARV S1133,ARV 1733,ARV 2408, ARV1017-1 M2 gene were 99.4%～99.6%, The homology of ARV C-98,ARV T-98 and ARV 176 M2 gene were 95.8%, The homology of ARV C-98,ARV T-98 and ARV OS161,ARV 918,ARV 916SI,ARV 601G,ARV138 M2 gene were 71.3%～73.3%,The homology of ARV C-98,ARV T-98 and DRV S14 M2 gene were 63.6%～63.7%, The low homology of ARV C-98,ARV T-98 and MRV T1L,MRV T3D M2 gene were 40.3%, respectively.The homology of ARV C-98,ARV T-98 and ARV S1133,ARV 1733,ARV 2408, ARV176 M3 gene were 98.3%～99.8%, The homology of ARV C-98,ARV T-98 and ARV 138,ARV OS161 M3 gene were 89.0%～92.3%, The homology of ARV C-98,ARV T-98 and ARV 918,ARV 1017-1,ARV 916SI M3 gene were 78.3%～89.3%,The homology of ARV C-98,ARV T-98 and DRV S14,DRV 89330 M3 gene were 67.7%～68.3%, The low homology of ARV C-98,ARV T-98 and MRV T1L,MRV T3D M3 gene were 21.5%～23.2%, respectively.