口蹄疫病毒复合多表位DNA疫苗的设计及构建

目的构建口蹄疫病毒(FMDV)复合多表位基因工程疫苗表达盒OAAT及其DNA疫苗。方法以O型、A型FMDV结构蛋白VP1全基因和Asia1型FMDV两个基因拓扑型的结构蛋白VP1基因上的5个抗原表位基因作为主要免疫原基因,非结构蛋白3ABC上的2个Th2细胞表位基因及结构蛋白VP4上的1个Th2细胞表位基因作为辅助基因,构建表达盒。将构建好的表达盒OAAT克隆到真核表达载体pVAX1 PCMV启动子下游,构建三价口蹄疫核酸疫苗pVAX1-OAAT,并用Western blot和IFA方法检测目的蛋白在HeLa细胞中的表达。结果通过InsightⅡ软件同源建模和DNAStar5.0软件分析表明,所构建的FMDV复合多表位基因工程疫苗表达盒OAAT理论上符合设计要求,且在真核细胞获得了正确表达。结论已成功设计FMDV复合多表位基因工程疫苗表达盒OAAT,并构建了三价口蹄疫核酸疫苗pVAX1-OAAT。

Design and Construction of Multiple-epitope DNA Vaccine against Foot-and-mouth Disease Virus

Objective To construct expression cassette OAAT for multiple-epitope antigen of foot-and-mouth disease virus(FMDV).Methods The cassette OAAT for multiple-epitope antigen of FMDV was constructed using the structural protein VP1 genes of FMDV serotypes A and O and 5 antigenic epitope genes of two genotypes of FMDV serotype Asial as major immunogen genes,and two Th2 epitope genes of non-structural protein 3ABC and one Th2 epitope gene of structural protein VP4 as accessory genes,then cloned downstream to eukaryotic expression vector pVAX1 PCMV.Transfect HeLa cells with the constructed trivalent FMDV DNA vaccine pVAX1-OAAT and identify the expressed product by Western blot and IFA.Results The modeling of homology by InsightⅡ software and analysis by DNAStar5.0 software proved that the design of constructed expression cassette OAAT was consistent with that exprected in theory.Western blot and IFA showed correct expression of target gene in Hela cells.Conclusion The expression cassette OAAT for multiple-epitope antigen of FMDV was successfully designed,and trivalent FMDV DNA vaccine pVAX1-OAAT was successfully constructed.