汉滩病毒西安分离84FLi的M片段全基因序列测定及分析

Sequencing and analyses of complete M segment sequence of 84FLi, a Hantaan virus isolated from Xi'an

目的 测定我国肾综合征出血热患者流产胎儿肝脏分离病毒 84 FL i株的全 M片段基因序列 ,了解其分子基础 .方法 采用反转录 -聚合酶链反应 (RT- PCR) ,分节段扩增 M基因片段的 c DNA,直接用 PCR产物或克隆入 p MD18- T载体后进行测序 .结果 84 FL i株的全 M基因序列组成为 :M片段基因共由 36 16个核苷酸组成 ,四种核苷酸的比例分别为A 30 .92 % ,C 18.0 3% ,G 2 1.18% ,T 2 9.87% . GC含量为39.2 1% ,AT含量为 6 0 .79% .最大开放读码框为 4 1～ 344 8,共编码 1135个氨基酸 .核苷酸序列同源性分析表明 84 FL i株与 HTN型同源性高于与其他型汉坦病毒的同源性 ,其中与中国株的同源性较高 ,与 HV114株的同源性为 85 .5 % .与HTNV的国际标准毒株 76 - 118的核苷酸同源性分别为84 .0 % ,氨基酸的同源性为 96 .0 % .结论 84 FL i株属于汉滩型病毒 。

AIM To study the complete M segment sequence of 84 FLi, a Hantaan virus isolated from a fetal liver in Xi'an and to explore its molecular characters. METHODS The cDNA of M segment was amplified fragment by fragment using RT PCR. The purified PCR products were sequenced directly or cloned into pMD18 T vector and then sequenced. RESULTS The complete M segment of 84FLi strain was composed of 3616 nucleotides with a single open reading frame,extending from 41 to 3448 encoding 1135 amino acids. Homology analysis showed that the homology of 84FLi M segment nucleotide sequences with HTN isolates, especially Chinese isolates, was higher than that of other Hantavarises, for example, 85.5% nucleotide sequences with HV114 (isolated in China), while only 84.0% with 76~118 (HTN foreign standard strain). The amino acid homology between the two strains was 96.0%. CONCLUSION The results suggest that Strain 84FLi is one of the Hantaan virus, which is highly related to other Chinese Hantaan virus isolates.