摘要
采用逆转录 聚合酶链式反应 (RT PCR) ,分节段扩增汉滩病毒 84FLi株的L基因片段cDNA ,纯化的PCR产物片段直接用于序列测定或克隆入 pND18 T载体中进行测序。结果表明 ,84FLi株的L片段cDNA由 6 5 33个核苷酸组成 ,四种核苷酸的比例分别为A33.39% ,C16 .4 3% ,G2 0 .74 % ,T2 9.4 4 %。GC含量为 37.17% ,AT含量为6 2 .83%。推导出的最大开放读码框架为从 38到 6 4 93,共编码 2 15 1个氨基酸。序列同源性分析表明 ,84FLi株核苷酸与HTN型国际标准毒株 76 118的同源性为 83.7% ,差异性为 18.8% ;而与中国株A9的同源性高达 97.6 % ,差异性仅为 2 .4 %。与SEO型代表Seoul80 39的同源性为 75 .2 % ,6 6 .1%～ 6 6 .5 %。L片段的氨基酸比较分析表明 ,L片段与HTN型间的同源性为 97.5 %～ 98.0 % ,而与SEO型的同源性为 83.5 %～ 85 .5 %。与PUC、TUL、SN和AND等其它型汉滩病毒的同源性仅为 6 8.6 %～ 6 9.6 %。结果表明 84FLi株属于HTN型 。
In order to study the nucleotide sequence of L segment of Hantaan virus strain 84FLi,the cDNA of L segment was amplified fragment by fragment by RT PCR.The purified PCR products were sequenced directly or cloned into pMD18 T vector and then sequenced.The L genome segment is 6533 nucleotides in length with a predicated region(from 38 to 6493) encoding 2151 amino acids.The entire sequence composition was A33.39%,C16.43%,G20.74%,T29.44%,and the GC and AT contents were 37.17 and 62.83%.Homology analysis showed that the homologies of 84FLi L segment nucleotide sequence with HTN isolates especially Chinese isolates were higher than those of other Hantavirus.It showed 83.7% homology with HTN foreign standard strain 76 118,while the amino acid homology between the two strains was 97.5%.The result suggested that 84FLi strain was one of the Hantann virus,and highly related to other Chinese Hantaan virus isolates.
出处
《中国病毒学》
CSCD
2002年第3期221-225,共5页
Virologica Sinica
