靶向UL5的siRNA对Ⅱ型单纯疱疹病毒抑制的实验分析研究.pdfVIP

靶向UL5的siRNA对Ⅱ型单纯疱疹病毒抑制的实验研究 摘 要 目的：应用m悄干扰技术研究靶向II型单纯疱疹病毒UL5基因的siRNA对病毒的 干扰作用。方法：在OeneBank上查找HSV-2UL5基因全序列，使用siRNA毛E线设计软 件对UL5的靶基因序列进行初步筛选；对候选靶mRNAJ挂行二级结构预测和热动力学 参数分析以及脱靶分析。用BLAST分析排除与其它基因有高度同源性的序列。本实验 从UL5基因中筛选出了五条序列作为靶序列并由靶序列分别设计合成出对应的正、反 2000脂质体转染HEK293细胞，转染 义寡核苷酸。siRNAJ恿过转染载体LipofeCtamine 12小时后收集并固定细胞，荧光显微镜下观察转染效果。收集转染48d,时后的细胞进 转染入HEK293细胞12小时后，收集细胞在荧光显微镜下可见荧光小点，而未转染 低上清液中病毒感染滴度，阴性对照组和siRNA374对病毒感染滴度无影响。结论：1、 可以特异性降低II型单纯疱疹病毒的复制水平。 关键词：HSV-2；U1-5；siRNA The of UL5 of Gene Virus StudyTargeting Herpes Simplextype 11withSmallInterferenceRNA Abstract Objective：Totheeffectofsmallinterference investigate UL5 of virus 11withRNAinterference geneherpes simplextype Methods：The ofUL5 genesequence wasfoundfromtheGeneBank．The targetgene theUL5 on wereselectedwithsiRNA software sequences design online，thesecondary structureofcandidatemRNA Was and target predicted，thethermodynamicparameters ofthemwere were with off-Target analyzed,thetargeted genesequencescompared BLASTtoemilinate with this anysequencessignificanthomology．Instudy,f如etarget wereselectedandsenseand sequences antisense oligonucleotides．weredesigned，their iRNAwastransfectedintoHEK293cellswith 2000in