腺病毒7型河南分离株全基因序列测定及重组分析

目的了解腺病毒7型河南分离株（hAdV35/Henan/2010）全基因特征及与其他腺病毒之间的进化重组关系。方法设计39对全长引物,采用RT-PCR方法扩增河南分离株腺病毒全序列,用DNASTAR中的SeqMan拼接全序,运用BioEdit进行序列剪齐,运用Mega4.0分析基因组编码蛋白hexon、fiber、E4和penton区域氨基酸与核苷酸的同源性,并与其他腺病毒序列绘制进化树,运用Simplot软件进行序列重组分析。结果 hAdV35/Henan/2010基因组全长35 234bp,与其他腺病毒相比,基因组氨基酸和核苷酸同源性为68.0%-99.8%和46.5%-99.5%。不同编码区hexon、fiber、E4和penton的核苷酸同源性分别为72.4%-95.4%、74.1%-97.0%、55.7%-92.7%和69.9%-99.3%。进化树分析显示河南分离株与腺病毒7型疫苗株同属一支,与腺病毒3型进化分支最近,Bootsacn分析显示与B亚属腺病毒在多个区域发生重组,尤其与腺病毒3在hexon区域高度重组。结论 hAdV35/Henan/2010与B族腺病毒亲缘关系较近,与hAdV3在hexon区域存在重组。

Genomic characteristics and recombination analysis of human adenovirus type 7 strains isolated from Henan Province,China

Objective To study the genomic characteristics of human adenovirus type 7（hAdV 7）strains isolated from Henan and the evolution and recombination of these strains with other adenovirus virus. Methods The entire genome sequence was amplified using reverse translation polymerase chain reaction（RT-PCR）with 39 pairs of primers.Sequences were spliced using the biological software SeqMan from DNAStar and sequences were aligned using Bioedit.The amino acid（aa）and nucleotide（nu）homology of the fiber,hexon,and E4 regions was analyzed and phylogenetic trees of the Henan strains and other adenovirus strains were drawn using Mega 4.0and recombination was analyzed with Simplot. Results The full length of the hAdV35/Henan/2010 genome was 35,234 bp.Compared to other adenoviruses,the Henan strains had aa similarity of 68.0%-99.8%and nu similarity of 46.5%-99.5%.The nu similarity of the hexon region was 72.4%-95.4%,that of the fiber region was 74.1%-97.0%,that of the E4 region was 55.7%-92.7%,and that of the penton region was 69.9%-99.3%.Phylogenetic analysis revealed that the Henan strains belonged to the same cluster as hAdV7 vaccine strains and had hAdV3 as their closest relative.Bootscan analysis revealed recombination with hAdV B strains in several regions,and particularly with hAdV3 in the hexon region. Conclusion hAdV35/Henan/2010 was closely related to hAdV B strains and had recombined with hAdV3 in the hexon region.