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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 To determine the whole gene sequence of Z10 vaccine against Haemorrhagic Fever with Renal Syndrome in China, understand its molecular structure and analyze the genetic differences between Z10 and other Hantavirus strains and provide basis for vaccine production. Methods Total RNA was extracted from cells infected with Z10 virus. The products amplified by reverse transcription polymerase chain reaction (RT PCR) were purified and cloned into T vector and sequenced. DNAS TAR software was used for analysis and comparison. The results of sequence analysis showed that the full-length sequences of L, M and S fragments of Z10 were composed of 655, 3366 and 170 nucleotides, respectively. The single-reading frame encoded 2115, 1135 and 429 Amino acids. The comparative analysis of genes showed that Z10 strain had lower homology with other Hantaan viruses except HTN strains and was similar to HTN virus but had only 83.6% to 87.4% homology with the HTN virus nucleotide sequence. , And its amino acid sequence homology as high as 94 2% ~ 96%, thus drawn three fragments of nucleotide and amino acid phylogenetic tree. Conclusion The whole genome sequence of Z10 strain was determined. The Z10 strain was the first reported subtype of HTN.