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浙江东方田鼠分离汉坦病毒株S基因的克隆和序列分析
Clonign and Sequence Analysis of S gene of hantavirus strain isolated from Microtus fortis In ZheJiang Province
【作者】 翁景清; 谢荣辉; 卢亦愚; 姚苹苹; 李敏红; 李婵; 徐芳; 茅海燕; 朱函坪; 崔庆荣; 董瑛; 朱智勇;
【Author】 Weng Jing-qing,Xie rong-hui,Lu yi-yu, et al. (Zhejiang Provincial Center for Disease Prevention and control,HangZhou 310009,China)
【机构】 浙江省疾病预防控制中心; 天台县疾病预防控制中心;
【摘要】 对国内首次从东方田鼠分离到的汉坦病毒ZT10,参考GenBank发表的的汉坦病毒核蛋白基因序列,设计合成一对引物,提取分离株ZT10细胞培养物的总RNA,对ZT10 S 基因进行RT-PCR扩增,产物经琼脂糖电泳分析,呈现一条约1.7kb的条带,回收纯化后, 将其克隆至pGEM-T栽体中,然后进行核苷酸序列的测定及分析。与四株SEO型病毒(SR-11、R22、Guo3、8610)核苷酸和氨基酸同源性分别为87.9%-96.0%和96.9%-97.9%, 与HTN型病毒株76-118的核苷酸和氨基酸同源性分别为71.3%和81.9%,与其它型汉坦病毒的同源均很低,表明此分离株ZT10为SEO型汉坦病毒,为进一步研究其进化和变异提供了有利条件,也为生产SEO型汉坦病毒特异性核蛋白抗原,免疫血清学诊断试剂的的制备和分子生物学研究奠定了基础.
【Abstract】 The S gene of hantavirus strain ZT10 isolated from M.fortis in ZheJiang Province was amplified by one step RT-PCR. The product of 1.7kb.fragment was obtained as expected. The amplified fragment was then cloned into the pGEM-T vector and confirmed by PCR and sequencing.The result demonstrate the S segment of ZT10 is composed of 1753 nuclotides in length with a open reading frame encoding a protein of 429 amino acids. Comparison with that of SEO type viruses (SR-11, R22, Guo3, 8610 strains) show 87.9%-96.0% homology at the nucleotide level, 96.9%-97.9% homology at the amino acid level respectively. Whik Comparison with HTN type virus (76-118 strain ) indicated that there was 72.0% at the nucleotide level, 82.6% homology at the amino acid level. Comparison with other type hantavirus show lower homology at the nucleotide and amino acid level respectively. The results of nucleotide and amino acid comparison indicated that ZT71 strain is SEO virus in molecular level.
【Key words】 Hantanvirus; ZT10 strain; S segment; Sequence analysis M.fortis;
- 【会议录名称】 2005年浙江省医学病毒学、医学微生物与免疫学学术会议论文汇编
- 【会议名称】2005年浙江省医学病毒学、医学微生物与免疫学学术会议
- 【会议时间】2005-07
- 【会议地点】中国浙江湖州
- 【分类号】R373
- 【主办单位】浙江省医学会医学病毒学分会、医学微生物与免疫学分会