Cloning And Sequence Analysis Of The S3, S4 Gene Of Avian Reovirus

The total RNA of Avian reovirus(ARV) isolated in inner Mongolia and Tianjin was extracted. According to the published S3 and S4 gene sequence of ARVs strains in GenBank,two pairs of primer were synthesized.The S3 and S4 gene of virus were amplificated with specific primers by RT-PCR and then cloned into PMD19-T plasmids and then sequenced. Each of the acquired sequences S3 gene was 1,202 bp,and contained one ORF(31-1,134 bp).Each of the acquired sequences S4 gene was 1,109 bp,and contained one ORF(2-1,105 bp). the accession nommber of ARV B-98,C-98,G-98,T-98 was EF030498,EF030496,EF030497,EF030499, respectively in GenBank.Comparision of nucleotide and amino acid sequences showed that high homology between ARVs.The homology of S3 gene was about 99.2%～100%,compared the four isolated ARV strains with ARV S1133,ARV 176,DRV YJL .The homology of S3 gene was about 87.0%～87.3%,compared the four isolated ARV strains with ARV 138. The homology of S3 gene was about 60.0%～64.1%,compared the four isolated ARV strains with DRV S14,DRV 89026.The low homology of S3 gene was 3.8% and 19.2%,respectively,compared the four isolated ARV strains with MRV 3 and NBV.The homology of S4 gene was about 98.8%～99.6%,compared the four isolated ARV strains with ARV S1133,ARV 176,DRV YJL .The homology of S4 gene was about 76.4%～81.2%,compared the four isolated ARV strains with ARV 138,DRV S14 and DRV 89026. The low homology of S4 gene was 6.5%～6.6% and 53.9%～56.2%, respectively, compared the four isolated ARV strains with MRV 3 and NBV.The results of phylogenetic tress indicated that the S3 and S4 gene of the four isolated ARV strains were close distant from ARV S1133,ARV 176 and DRV YJL.The Structural analysis ofσB protein andσNS protein indicated that they were hydropathicity protein,and isoelectric point were 6.27～7.10. The antigen site ofσB protein andσNS protein of four ARV isolates were similar with ARV S1133,ARV 176 and DRV YJL.The analysis results of homology,phylogenetic tress and antigen site were identical.