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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 perform genome-wide sequencing of the adaptive strain of tick-borne encephalitis virus (strain TBZ) in Vero cells (hereinafter referred to as SZV strain) and to compare with other strains of Far East subtype of TBEV , European subtype strains were compared. Methods The specific primers of TBEV were designed and the total RNA of SZV strain was extracted. The amplified fragment was sequenced by RT-PCR and sequenced. Vector NTI 9.0 software was used for splicing. , GenBank TBEV far-eastern-type Oshima 5-10 strains, Sofjin-HO strains and European subtype 263 strains, Neudoerfl strains of the full-length gene sequences for comparison analysis. Results The whole genome of SZV strain was composed of 10 782 nucleotides, encoding only 1 open reading frame (ORF) of 10 245 nucleotides in length and encoding a total of 3 414 amino acids. The amino acid sequence identities of SZV strain and ORF region of Far East Asian TBEV strain were 95.0% and 94.6%, respectively, which were close to those of typical strains of TBEV European subtype strain 263 and Neudoerfl strain ORF region The encoded amino acid sequence homologies were 91.1% and 91.0%, respectively. Conclusion Genome sequencing of the SZV strain of Vero cell adapted to TBEV and its analysis have laid the foundation for the development of Vero cell-based vaccine for forest encephalitis.