森林脑炎病毒Vero细胞适应株的全基因组序列分析  

Whole genome sequencing of Vero cell-adapted tick-borne encephalitis virus strain

作　　者：孙宏亮[1] 王伟[1] 刘双军[1] 麻红梅[1] 张朔[1] 邢坤鹏 李野[1] 王涛[1] 宋宗明[1] 

机构地区：[1]长春生物制品研究所有限责任公司疫苗三室,吉林长春130062

出　　处：《中国生物制品学杂志》2015年第6期553-556,共4页Chinese Journal of Biologicals

基　　金：国家科技部重大新药创制项目(2013ZX09402302-205)

摘　　要：目的对森林脑炎病毒(tick-borne encephalitis virus,TBEV)森"张"株在Vero细胞上的适应株(以下称SZV株)进行全基因组测序,并与TBEV远东亚型的其他毒株、欧洲亚型毒株进行比较。方法设计TBEV的特异性引物,提取SZV株病毒总RNA,以其为模板,采用RT-PCR方法分段扩增序列并测序,应用Vector NTI 9.0软件进行拼接,并与森"张"株、Gen Bank中报道的TBEV远东亚型Oshima 5-10株、Sofjin-HO株及欧洲亚型263株、Neudoerfl株的全长基因序列进行比对分析。结果 SZV株的全基因组由10 782个核苷酸组成,仅编码1个开放阅读框,长度为10 245个核苷酸,共编码3 414个氨基酸。SZV株与TBEV远东亚型的Oshima5-10株和Sofjin-HO株ORF区编码的氨基酸序列同源性分别为95.0%和94.6%,与TBEV欧洲亚型的典型毒株263株和Neudoerfl株ORF区编码的氨基酸序列同源性分别为91.1%和91.0%。结论对TBEV Vero细胞适应株SZV进行了全基因组序列测定及分析,为研制基于Vero细胞基质的森林脑炎疫苗奠定了基础。Objective To sequence the whole genome of Vero cell-adapted tick-borne encephalitis virus（TBEV） SenZhang strain（SZV strain） and compare with those of other TBEV strains of Far East and Europe subtypes. Methods Specific primers for TBEV were designed, with which the total RNA was extracted from SZV strain and used as a template for RT-PCR. The amplified products were sequenced and spliced by using Vector NTI 9. 0 software, then compared with the full-length gene sequences of SZV as well as Oshima 5-10 and Sofjin-HO strains of Far East subtype and 263 and Neudoerfl strains of Europe subtype in Gen Bank. Results The whole genome of SZV strain consisted of 10 782 nucleotides, which encoded a single open reading frame（ORF） at a length of 10 245 nucleotides encoding 3 414 amino acids. The homologies of amino acid sequences of SZV strain were 95. 0% and 94. 6% to the ORFs of Oshima 5-10 and Sofjin-HO strains, and 91. 1% and 91. 0% to those of 263 and Neudoerfl strains, respectively. Conclusion The whole genome of SZV strain was sequenced, which laid a foundation of preparation of Vero cells-derived tick-borne encephalitis vaccine.

关 键 词：森林脑炎病毒 VERO细胞 序列分析 

分 类 号：R373.31[医药卫生—病原生物学] Q71[医药卫生—基础医学]