文档介绍：甘肃农业大学硕士学位论文 AsiaⅠ型口蹄疫基因治疗与基因免疫双功能疫苗载体的构建姓名:项海涛申请学位级别:硕士专业:预防兽医学指导教师:柳纪省;胡永浩 20090601 甘肃农业大学2009屑硕十学位论文 AsiaI型口蹄疫基因治疗与基因免疫双功能疫苗载体的构建摘要目前,对于口蹄疫的防制主要采用屠杀、隔离和免疫接种,其中疾病流行前的免疫接种是特异性保护家畜免遭感染的有效途径,但现有疫苗接种动物后都会存在免疫空白期,即诱导免疫动物产生体液和细胞免疫反应,等待有效抗体产生的一段时间。动物如果在这段时间内遭到病毒的入侵往往会引起该病的流行。为探索解决免疫动物在疫苗接种的免疫空白期内发病这一难题,本研究以Asia I 江苏株为模板,引入RNA干扰机制,进行了双功能疫苗的探索研究,并取得下述进展: ,将其CMV启动子替换为U6启动子,。 ,结果表明对载体的改造是成功的, 的两个启动子均能启动目的基因的表达。 I江苏株的3D基因设计3条小RNA干扰片段,并将之与免疫组合基因P12A+,成功构建出了3个双功能疫苗质粒。 ,有延迟细胞病变的作用,说明双功能载体能够转录出siRNA,对病毒的转录复制有一定的抑制效果。 。关键词:双功能疫苗载体;FMDV;RNAi; 甘肃农业大学2009屑硕十学位论文,1 』●●■ o』I - LOnstruCtlon 01 aDi-IUnctional lne vector ot disease virus Summary Atpresent,slaughter,isolation ination aremajor methods tocontrolFMD, and ination before epidemic ofFMD isthe specificefficient method toprotect animals against achallenge tothedevelopment of ines against microbiology isthe existofimmunization black and the latentpedod,FMD would outbreak when animalschallenged wim virusduring theimmunization black aswell as animals ine during order tosolvethis problem,we introducedRNAi intothebi-functional ine research,using AsiaI/JS strain asthemodel toconfirmthefeasibility oftheconception. theCMV promoter of vector by U6promoter,developed adual-promoter vector . thefunctionsofthepromoters byexpressing EGFP and Interfering EGFP de呻nstrated thatbothCMV promoter锄d U6 promoter could play constructedthreebi—functionalplasmids expressing shRNAs targeting 3D gene ofAsiaI/JSstrainFMDV andexpressing immunization protein P 2A+3 Csimultaneously. antiviralpotential induced by bi-functionalplasmid Was evidentinthe face of challenge of with a homologous FMD