汉滩病毒Z10株G1糖蛋白编码区克隆及序列分析

Molecular cloning and nucleotides sequence analysis of G1 genome segment of Hantaan virus Z10 strain

目的 汉滩病毒浙 10 (Z10 )株G1糖蛋白编码区的克隆、核苷酸序列分析 ,提供我国应用最为广泛的肾综合征出血热 (HFRS)疫苗株序列资料。方法 反转录PCR法扩增G1基因片段 ,克隆入pGEM T载体 ,双脱氧链终止法测定核苷酸序列。结果 测定 144 9个核苷酸 ,可编码 483个氨基酸。与Ⅰ型 76 118、Lee、Hojo株比较核苷酸同源性分别为 87%、86 %、86 % ,与Ⅱ型R2 2株同源性为6 7%。比较氨基酸序列Z10株与Ⅰ型、Ⅱ型病毒的同源性分别为 (94～ 95 ) % ,(77～ 80 ) %。Z10与 76 118G1编码的氨基酸变异多发生于N′端 ,最大的变异区是第 84～ 93位氨基酸的 10个连续变异。结论Z10株病毒和其它汉滩病毒在核苷酸序列虽有较大差异 ,但在氨基酸水平上的同源性仍较高 ,可能是Z10株沙鼠肾疫苗具有较好免疫保护作用的原因。

Objective To study the sequence of HFRS vaccine strain that is widely used in China. Methods The G1 genome segment of Z10 virus was amplified with RT PCR, and after being identified and purified the products were cloned into pGEM T vector. The clone was sequenced by Sanger's dideoxy chain termination method. Results 1?449 bases of Z10 G1 genome segment, coding for 483 amino acids were determined. The base compositions for the G1 segment RNA determined from cDNA sequence information were 30.8% A, 21.9% G, 19.1% C and 28.2% U. These values were similar to those of the 76/118, Lee and SR virus. As compared to Z10 G1 segment, the sequence homology at the nucleotide level was 87%(76/118, type Ⅰ), 86% (Lee, type Ⅰ),86% (Hojo, type Ⅰ), 67%(R22,type Ⅱ), and 59%(K22, type Ⅲ).The Z10 virus G1 protein had higher amino acid sequence identity with other Hantaan viruses (94% 95% homology) than Seoul type viruses (77% 80%). More amino acid sequence heterogeneity between the Z10 and 76/118 was observed in N terminal, especially cluster of divergence of ten amino acids at position 84 to 93 of the Z10 virus G1 protein. Conclusions (1) Z10 strain is one of the Hantaan viruses. (2) The important region of G1 segment of Z10 was conservative. (3) Although substantial divergence of nucleotide sequences of the G1 genome segment was found between the Z10 strain and other type I Hantaan viruses, relatively high amino acid sequence homology was shown among them. Thus, good immune protection could be obtained with the inactivated vaccine grown on Meriones unguiculatus kidney cells against other strains of Hantaan viruses.