汉坦病毒Z10株M片段全基因序列分析

Complete sequence analysis of the M genome segment of Hantavirus Z10 strain

目的 测定我国疫苗株Z10的M片段全基因序列 ,了解其分子基础 ,并分析其与其它汉坦病毒株的遗传学差异。方法 从Z10株病毒感染的细胞提取总RNA ,将逆转录PCR扩增的产物纯化后克隆于T载体并进行序列测定 ,应用DNASTAR软件分析比较。结果 Z10株M片段的全基因序列共 36 15个核苷酸 ,四种核苷酸的比例分别为A 30 15 % ,T 2 9 41% ,G 2 1 72 % ,C 18 73%。以第二种读码方式推导出其最大读码框架从 41到 34 48,共编码 1135个氨基酸。序列同源分析表明 ,Z10株与HTN型核苷酸同源性为 83 6 %～ 87 4% ,与SEO ,DOB和Thai型为 6 9 5 %～ 70 8% ,而与PUU和HPS各型病毒同源性仅为 5 7 6 %～ 5 9 9%。氨基酸同源分析Z10株与HTN型同源性高达 94 2 %～96 0 % ,与SEO ,DOB和Thai型为 73 3%～ 76 6 % ,而与PUU和HPS各型病毒同源仅为 5 3 2 %～5 6 3%。绘出了核苷酸和氨基酸的系统发生树。结论 Z10株为不同于已报告M片段全基因序列的HTN型国内外毒株的另一亚型 ,并具有 16个独特的氨基酸。

Objective To Sequence the complete sequence of the M genome of Hantavirus Z10 strain which has been exploited for inactivated vaccine production in China, to explore its molecular characteristics and compare it with other Hantaviruses. Methods The total RNA were prepared from Z10 virus infected cells and the RT PCR product was cloned into T vector, sequenced and analyzed using DNASTAR software. Results The Z10 complete M segment was 3615 nucleotides in length with a single open reading frame extending from 41 to 3448 encoding 1135 amino acids. It showed 83.6% 87.4% homology with other 7 HTN viruses at the nucleotide level and 94.2% 96% at the amino acid level, only 69.5% 70.8% homologous with SEO, DOB & Thai viruses at the nucleotide level and 73.3% 76.6% at the amino acid level, while just 57.6% 59.9% homologous with PUU, HPS viruses at the nucleotide level and 53.2% 56.3% at the amino acid level. We got the phylogenetic tree depending on the nucleotide and amino acid sequences. Conclusion The results suggest that Z10 strain is a new subtype with 16 unique amino acids different from other HTN viruses