【摘要】 目的对国内首次从东方田鼠分离到的汉坦病毒(HV)ZT10S基因克隆进行序列分析。方法参考GenBank发表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ZT10S基因进行RT-PCR扩增,产物经琼脂糖电泳分析,呈现一条约1.7 kb的条带,回收纯化后,将其克隆至pGEM-T载体中,然后进行核苷酸序列的测定及分析。结果与4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%,与其他型汉坦病毒的同源性均很低,表明此分离株ZT10为SEO型汉坦病毒。结论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制备和分子生物学研究奠定了基础。更多还原

【Abstract】 Objective The S gene of hantavirus strain ZT10 isolated from M.fortis in Zhejiang province was amplified by one step RT-PCR.Methods The product of 1.7 kb fragment was obtained as expected.The amplified fragment was then cloned into the pGEM-T vector and confirmed by PCR and sequencing.Results The result demonstrates that 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) shows 87.9%-96.0% homology at the nucleotide level,96.9%-97.9% homology at the amino acid level respectively.Comparison with HTN type virus(76-118 strain) indicated that there was 72.0% homology 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.Conclusion The results of nucleotide and amino acid comparison indicated that ZT71 strain is SEO virus in molecular level.更多还原