西尼罗病毒Chin-01株基因组非编码区的序列测定及分析  

Determination and analysis of the sequence of untranslated region of West Nile virus Chin-01 RNA

作　　者：李晓峰[1] 姜涛[1] 陈水平[1] 韩剑峰[1] 邓永强[1] 秦成峰[1] 于曼[1] 秦鄂德[1] 

机构地区：[1]军事医学科学院微生物流行病研究所、病原微生物生物安全国家重点实验室,北京100071

出　　处：《解放军医学杂志》2008年第7期829-832,共4页Medical Journal of Chinese People's Liberation Army

基　　金：国家“973”计划资助项目(2002CB513205)

摘　　要：目的对西尼罗病毒(WNV)Chin-01株基因组非编码区(UTR)序列进行测定,了解其与已报道毒株的序列差异及系统发育关系。方法采用5′和3′RACE法对Chin-01株病毒基因组的UTR进行RT-PCR扩增,测定扩增产物序列,采用DNAstar软件对该株及其他9株WNV的UTR进行序列同源性分析,同时构建了基于3′UTR的系统发育树,并用RNA structure软件预测其二级结构。结果Chin-01株基因组5′和3′UTR分别为96nt和630nt,序列同源性分析的结果表明,其5′UTR与其他9株的同源性均达到99%或100%,而3′区UTR与埃及Eg101株的同源性最高,达98·1%。5′UTR的起始核苷酸为AG,并含有一段与3′UTR互补的14nt序列。3′UTR的末端为CUOH,包含一段完全保守的5nt序列,以及三个保守的称为CS2、RCS2和CS1的基序。二级结构预测发现,Chin-01株5′UTR呈长茎环结构,3′UTR的3′端也可形成多个保守的茎环结构。结论Chin-01株与埃及Eg101株亲缘关系最为密切。Objectives To study the interrelation and variation of the untranslated regions（UTR） between West Nile virus（WNV） strain Chin-01 and other WNV strains and lay a foundation to study the functions of the UTR by sequencing and analyzing the sequence of UTR of WNV strain Chin-01.Methods Using the 5＇and 3＇RACE,the sequences of 5＇and 3＇UTR of strain Chin-01 were amplified by reverse transcription polymerase chain reaction（RT-PCR）,and the cDNA fragments obtained were sequenced and then analyzed with DNAstar software,the secondary structures of which were also predicted with RNA structure software.In addition,the phylogenetic tree based on the 3＇UTR was constructed.Results The sequences of 5＇ and 3＇ UTR of strain Chin-01 contain 96 nucleotides（nt） and 630 nt,respectively.Sequence homology analysis revealed that the 5＇ UTR showed a sequence identity of 99% or 100% among 10 strains of WNV,while the 3＇ UTR exhibited the highest sequence identity of 98.1% with strain Eg101.5＇terminal nucleotides of the 5＇UTR end as 5＇AG,in which there is a putative sequence of 14 nt complementary to a sequence located in the 3＇UTR.The 3＇UTR terminates as CUOH-3＇ and contains a completely conserved sequence of 5 nt and three conserved sequences named CS1,CS2 and RCS2.The prediction of secondary structure revealed that the structure of 5＇UTR consisted of a stem with a small top loop and a larger side loop.The 3＇UTR also contained several conserved secondary structures.Conclution There is a closest evolutionary relationship between the Chin-01 strain and Eg101 strain,both of which belong to the lineage I.

关 键 词：西尼罗病毒 非翻译区 序列分析 蛋白质结构 二级 

分 类 号：R373.33[医药卫生—病原生物学] R512.91[医药卫生—基础医学]