A组轮状病毒广州地方株VP7基因序列的比较分析

Sequence analysis of VP7 gene from rotavirus field strain from Guangzhou, China

目的 了解我国轮状病毒G1型广州地方株与标准株及北京G1型地方株VP7基因序列的差异 ,为我国轮状病毒疫苗的研制提供资料。方法 通过逆转录 聚合酶链反应 (RT PCR)获得了轮状病毒广州地方株R97 196VP7全基因的cDNA片段 ,将其克隆入T A克隆质粒pUCm T中 ,构建成重组质粒pUCmT VP7,对克隆的VP7基因进行序列测定。结果 该地方株的基因核苷酸全长为10 6 2nt,读码框架和以往的研究一致 ,和北京G1型地方株T73的VP7氨基酸序列具有高度同源性(98% ) ,而与不同血清型标准株间则变异较大 (73%～ 81% ) ,氨基酸序列中存在的一些高变区和保守功能区与已报道的研究结果一致。从进化角度分析 ,与轮状病毒标准株Wa株 ,相距较远。结论 轮状病毒广州地方株R97 196VP7基因片段属G1型 ,轮状病毒VP7基因的变异与地域有一定关系。

Objective To compare the difference of VP7 fragment at the nucleotide level between rotavirus Guangzhou field strain R97 196 and rotavirus Beijing field strain or prototype strains Methods The VP7 fragment amplified from Guangzhou field strain R97 196 by RT PCR was cloned into the T A cloning vector pUCm T and sequenced Results The VP7 fragment from Guangzhou field strain R97 196 was 1 062 bp in length and contained two open reading frames which is consistent with that reported in the literature The sequence analysis revealed that the cDNA from R97 196 shared higher nucleotide and amino acid identities with Beijing field strain T73 (98% and 98%, respectively) than with serotype 2 4 ( G types ) rotavirus (from 74% to 77% and 73% to 81%, respectively) The divergence of amino acid sequences is mainly within the nine divergence regions The Phylogenetic analysis revealed that the VP7 fragment from R97 196 was far away from rotavirus serotype G1 strain Wa Conclusion The VP7 gene fragment from rotavirus Guangzhou field strain R97 196 belongs to rotavirus serotype G1 And variation of rotavirus VP7 gene fragment seems to be a geographic matter