【摘要】 以pcDNA3质粒为骨架,构建由1型单纯疱疹病毒(Herpes simplex virus I,HSVI)立即早期基因启动子指导外源蛋白转录的phIE真核表达载体,该质粒为外源蛋白的表达提供了更为广泛的选择,通过插入us1编码序列,phIE-Us1被用于分析ICP22蛋白在病毒感染早期的功能;在多克隆位点插入氯霉素乙酰转移酶编码基因构建phIE-CAT检测载体,应用于检测HSVI病毒蛋白Vp16及ICP22的功能,明确显示该质粒对于研究单纯疱疹病毒的基因转录调控具有重要的应用价值。更多还原

【Abstract】 An eukaryotic expression vector phIE composed of pcDNA3 skeleton and the herpes simplex virus immediate early gene promoter was constructed to provide a new choice for ectopic expression.The plasmid phIE-Us1 was constructed by inserting the us1 cDNA downstream of the promoter to analyze the function of ICP22 protein very early in the infection process.Furthermore,the gene encoding the chloramphenicol acetyl-transferase was inserted into the multiple cloning site of the expression vector to yield the phIE-CAT.In functional analysis of viral protein ICP22 and Vp16,phIE-CAT was confirmed to be capable of evaluating the mechanism of the genome of herpes simplex virus 1 transcriptional regulation.更多还原