汉坦病毒Z10株全基因序列的测定及分析

Complete genome sequence analysis of the Hantavirus Z10 strain

目的 测定我国肾综合征出血热疫苗株Z10的全基因序列 ,了解其分子结构特征 ,并分析Z10与其他汉坦病毒株的遗传学差异 ,为疫苗生产提供依据。方法 从Z10病毒感染的细胞提取总RNA ,将逆转录 聚合酶链反应 (RT PCR)扩增的产物纯化后克隆于T载体并进行序列测定 ,应用DNAS TAR软件分析比较。结果 序列测定表明 ,Z10株L ,M ,S片段的全基因序列分别由 6 5 5 3,36 15 ,170 1个核苷酸组成 ,单一读码框架分别编码 2 15 1,1135 ,42 9个氨基酸。基因比较分析表明 ,Z10株与HTN型外其他汉坦病毒同源性较低 ,与HTN型病毒接近 ,但与HTN型病毒核苷酸序列的同源性亦仅为 83 6 %～ 87 4% ,而其氨基酸序列的同源性高达 94 2 %～ 96 % ,据此绘出了三个片段的核苷酸和氨基酸的种系发生树。结论 测定了Z10株的全基因组序列 ,首次发现Z10株为已报道的HTN型中的另一亚型。

Objective Study on the complete genome sequence of Hantavirus Z10 strain which has been applied for inactivated vaccine production in China,to assess its molecular characteristics and the diversity with oth er hantaviruses.Methods The total RNA were prepared from Z10 virus infected cells and the RT PCR products was cloned into T vector,sequenced and analyzed by using DNASTAR software.Results The Z10 complete genome,L segment is 6 553,M segment is 3 615,S segment is 1 701 nucleotides in length,with a single open reading frame encoding 2 151,1 135,429 amino acids respectively.Sequence homology comparison showed that the 3 segment nucleotide of Z10 strain were close to HTN type virus,but only 83 6～87 4% homology with other HTN viruses at the nucleotide level.The phylogenetic analysis was made on their nucleotide and amino acid sequences.Conclusion The results firstly demonstrates that Z10 strain is a new subtype of the Hantaan(HTN) type.