真核表达东方马脑炎病毒衣壳蛋白Capsid抑制IFNγ应答基因转录  

EEEV capsid inhibits transcription of IFNγ stimulated gene in 293 cell lines

作　　者：崔艳[1] 刘海楠[1] 李伟[1] 李平[1] 曹诚[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850

出　　处：《军事医学》2012年第7期512-515,共4页Military Medical Sciences

摘　　要：目的克隆东方马脑炎病毒(EEEV)衣壳蛋白(Capsid)基因,构建其真核表达载体,检测该基因在293细胞中的表达,并初步研究其对干扰素(IFN)应答基因表达的影响。方法构建包含EEEV capsid基因的真核表达载体pcDNA3-Flag-capsid,以威格拉斯脂质体转染293细胞进行瞬时表达,Western印迹检测其表达情况;应用定量PCR比较IFN效应基因Gbp1,Gbp2和Isg15在pcDNA3-Flag-capsid和空载体转染细胞中mRNA表达差异。结果构建了EEEV Capsid真核表达质粒,并且该质粒能够在293细胞中有效表达,同时定量PCR结果表明,与空载体转染对照组相比,外源导入EEEV Capsid抑制IFN应答基因Gbp1,Gbp2和Isg15的转录。结论真核表达EEEVCapsid抑制细胞内IFN应答基因的表达,为研究EEEV致病机制奠定基础。Objective To construct a eukaryotic expression vector of eastern equine encephalitis virus （EEEV） capsid genes, and to investigate the possibility of capsid involvement in the regulation of the expression level of the IFN stimulated gene. Methods The recombinant expression vector containing the capsid gene of EEEV was transiently transfected into 293 cell lines by VigoFect reagent. The QPCR was used to evaluate the effect of the EEEV eapsid on the transcription of IFN stimulated genes, such as Gbp1, Gbp2 and IsglS. Results The eukaryotic expression plasmid pcDNA3-Flag-Capsid was constructed and successfully expressed in 293 cell lines. The mRNA level of Gbpl, Gbp2 and Isg15 induced by INFγ was decreased in the EEEV eapsid expressed cells compared to the control. Conclusion The IFN stimulated gene expression is inhibited by the EEEV capsid, facilitating the research on the pathogenic mechanism of EEEV.

关 键 词：马脑炎病毒 东方 衣壳蛋白质类 真核表达 IFN应答基因 

分 类 号：R373.31[医药卫生—病原生物学]