肾综合征出血热灭活疫苗毒株L99株的全基因序列分析

Sequence Analysis of Whole Gene of Strain L99 for Preparation of Inactivated EHF Vaccine

目的 了解我国肾综合征出血热疫苗生产毒株Ｌ９９株的分子特征及抗原性的分子基础。方法 设计出不同的引物，用ＰＣＲ方法提取细胞总ＲＮＡ，逆转录扩增产物纯化后克隆Ｔ载体，纯化后测序，序列用ＤＮＡＳ ＴＡＲ软件拼接，并对其序列进行分析。结果Ｌ９９株全基因组由Ｌ片段６５３３个，Ｍ３６５２个，Ｓ１７６４个核苷酸组成， 分别编码２１５１、１１３３、４２９个氨基酸，Ｌ９９与同型病毒间同源率高达９５．５％－９９．４％。结论 为研究疫苗毒株的生物 学特性、致病机理等提供了分子基础。

Objective To analyze the sequence of whole gene and study on the molecular mechanism of epidemic hemorrhagic fever( EHF) virus strain L99 for preparing inactivated vaccine. Methods The total RNA of infected cells was extracted and amplified by RT-PCR using designed primers, then purified and cloned to T-vector. The recombinant plasmid was purified and sequenced, and the sequence of it was analyzed by DNASTAR software and compared with that of virus strains of other SEO types. Results The whole gene of strain L99 consisted of fragments L(6533bp) , M(3652bp)and S( 1764bp) coding 2151,1133 and 429 ammo acids respectively, and the homology of it to those of viruses of other SEO types reached 95.5%- 99.4%. Conclusion This paper provided a molecular basis of study on the biological characteristic and pathogenic mechanism of strain L99.