森林脑炎病毒森张株5′端非编码区序列测定

Nucleotide sequence of 5′-untranslated region of the tick-borne encephalitis virus Senzhang strain isolated in China

为了解森林脑炎病毒森张株 5′端非编码区序列与生物特性的关系 ,从而为疫苗研制打下基础 ,对其核苷酸序列进行测定。采用RACE法进行RT PCR扩增 ,克隆法测序。结果表明 :森张株 5′ UTR长 12 9nt,与Sofjin HO、Os hima5 10、Vasilchenko、Neudoerfl、2 6 3、Hypr的同源性分别为 92 %、91%、89%、87%、87%、87%。森张株存在三处特异性变异C18→T、T3 0 →C及 4 9、5 0连续两个核苷酸缺失。本实验建立了一套相似 5′端非编码区序列测定方法 ;三处特异性变异可能影响到森林脑炎病毒森张株的转录。

To understand the relation between TBEV 5′-untranslated region(5′-UTR) sequence and biological function and aid in the development of new Tick-borne Encephalitis (TBE)vaccines,the nucleotide sequence of Senzhang strain5′-UTR was determined.By RACE method,cDNA fragment of Senzhang strain 5′-UTR were acquired from infected BHK-21 cells.Then the cDNA fragments was cloned into the vector pGEM R-T and transformed to competent DH5α cells.positive clones were screened and identied by PCR.The sequences of inserted fragment were determented by 3100Genetic Anaiyzer.The results showed the5′-UTR of Senzhang strain containted 129 nucleotides(nt).The nucleotides sequence of Senzhang strain were compared with TBEV strains such as Sofjin-HO?Oshima5-10?Vasilchenko?Neudoerfl?263 and Hypr, the results revealed that the homogeneity among the seven strains was 92%?91%?89%?87%?87%and87%respectively,Senzhang strain had three specific varlations;C 18→T,T 30→C and diletion in 49-50nt.The method about how to determine the similar virus5′-UTR sequence was found.The three variations may affect on transcribtion,translation and encapsidation of Senshang strin.