E基因在辛德毕斯病毒与辛德毕斯样病毒细胞感染特性中的作用

目的 寻找辛德毕斯病毒(YN87448病毒)与辛德毕斯样病毒(XJ-160病毒)对细胞感染特性差异的分子基础.方法 生物信息学比较两种病毒糖蛋白基因一级结构及二级结构,分析二者之间的差异,并利用病毒基因重组等现代分子生物学技术构建重组病毒来研究糖蛋白基因在病毒感染细胞特性中的贡献.结果 生物信息学分析显示,YN87448病毒和XJ-160病毒基因组分别具有11 717和11 626核苷酸序列,具有相同的基因组结构特征;两种病毒E基因氨基酸的疏水性主峰基本相同但存在82个散在分布的氨基酸差异位点.病毒基因重组研究结果显示,将XJ-160病毒E基因替换为YN87448病毒E基因的重组病毒(XJ-160/YE1E2)无论在致细胞病变时间,空斑形成直径,还是在病毒功能蛋白的表达等方面均完全体现YN87448病毒特征,而与XJ-160病毒野毒株的表现完全不同.结论 E基因在辛德毕斯病毒与辛德毕斯样病毒细胞感染特性差异方面起着重要作用,这一结果为从分子生物学角度解释两病毒间生物学差异提供了分子生物学理论依据.

Role of the E gene in the infectivity of SINV and SINL in infected cells

Objective To elucidate the molecular basis on the differences of infectivity in infected cells between Sindbis virus(SINV:YN87448 virus)and Sindbis like virus(SINLV:XJ-160 virus).Methods Compare the E(glycoprotein)gene sequence and secondary structure of YN87448 virus and XJ-160 virus by bioinformatics analysis.Analyze the contribution of E gene to the biological differences between SINV and SINLV by constructing recombinant virus.Results By bioinformatics analysis,YN87448 virus and XJ-160 virus have the same genomic structure,which has 11 717 nt and 11 626 nt respectively.There are 82 amino acid differences between E gene of these two viruses,and showed scattered distribution.The main peak is basically the same for the hydrophobic of the E gene protein,but in some region existing small differences.The recombinant virus which exchanged the E gene of XJ-160 virus with YN87448 virus totally showed the biological character of YN87448 virus,either in the showing time of CPE,plaque forming time and plaque diameter,or in expression of functional proteins.Conclusion E gene plays a major role in the differences of infectivity in infected cells between SINV and SINLV,this result provide the molecular biological evidences for elucidating the biological differences between SINV and SINLV.