摘要
目的 克隆和测定汉滩病毒疫苗生产株 84FLi的全基因组序列 ,了解其分子基础。方法 采用逆转录聚合酶链反应 (RT PCR) ,分节段扩增 84FLi株L、M和S基因片段的cDNA ,直接用PCR产物或克隆入pMD18 T载体后进行测序。结果 84FLi株L、M、S 3个片段的全基因组序列为6 5 33,36 16和 16 88个核苷酸 ,分别编码 2 15 1,1135 ,4 2 9个氨基酸。A、C、G、T 4种核苷酸分别为 3830 ,2 0 5 0 ,2 5 10和 344 7个 ,GC含量为 38 5 2 % ,AT含量为 6 1 4 8%。序列同源性分析表明 84FLi株与分离自广州的RG9株和分离自安徽的Chen4株高度同源。S片段核苷酸的同源性 99 6 %。与HTNV的国际标准毒株 76 118的 3个片段核苷酸同源性分别为 83 7%、84 0 %和 87 2 % ,氨基酸同源性分别为97 5 %、96 0 %和 97 9%。结论 84FLi株属于汉滩型病毒 ,并与分离自国内的汉滩病毒同源性较高 。
Objective To study the complete genome sequence of the Chinese Hantaan virus vaccine strain 84FLi and learn about its molecular characters Methods The virus strain 84FLi was isolated from the liver of a fetus aborted by pregnant women with hemorrhagic fever with renal syndrome The cDNAs of L,M and S segments were amplified fragment by fragment using RT PCR The purified PCR products were sequenced directly or cloned into pMD18 T vector and then sequenced Results The complete genome of strain 84FLi was composed of L (6 533 bp), M (3 616 bp) and S (1 688 bp), coding 2151,1135 and 429 amino acids,respectively The entire sequence composition was 3830A,2050C,2510G and 3447T,the GC and AT contents were 38 52% and 61 48%. Homology analysis showed that the homologies of 84FLi S segment nucleotide sequences with strain RG9 (isolated in Guangzhou) and strain Chen4 (isolated in Anhui) were 99 6% There were 83 7%,84 0% and 87 2% nucleotide sequences homology with the three segments of Hantaan virus foreign standard strain-76 118, while the amino acids sequences homology with those of 76 118 were 97 5%,96 0% and 97 9% respectively Conclusion The strain 84FLi is highly related to other Chinese Hantaan virus isolates and is in the same subtype with the other two Chinese isolates RG9 and Chen4
出处
《中华医学杂志》
CAS
CSCD
北大核心
2002年第14期981-985,共5页
National Medical Journal of China
