PEI磁珠吸附IBRV主要囊膜蛋白基因的DNA疫苗的构建  

Construction of IBRV DNA Vaccine by PEI Absorbed the Tegument Protein Genes

作　　者：于国伟 刘行波[1] 倪宏波[1] Yu Guowei;Liu Xingbo;Ni Hongbo(College of Animal Science and Technology,Heilongjiang A滋gust First Land Reclamation University,Daqing 163319)

机构地区：[1]黑龙江八一农垦大学动物科技学院,大庆163319

出　　处：《黑龙江八一农垦大学学报》2020年第4期15-19,46,共6页journal of heilongjiang bayi agricultural university

基　　金：中央引导地方科技发展专项项目(ZY17C08)。

摘　　要：通过利用实验已经构建好的表达IBRVgB/gC/gD囊膜蛋白的真核表达质粒(pCDNA4-gB-His、pCDNA4-gC-His、pCDNA4-gD-His)为免疫原,以PEI磁珠为载体,PEG为保护层构建DNA疫苗。将构建好的真核表达质粒,通过鉴定、扩增、大提后,使PEI磁珠与其吸附并转染至小鼠胚胎细胞(NIH/3T3)鉴定蛋白表达情况,此后进行疫苗制备并通过电镜观察其形态特征。结果表明:三种蛋白均能在小鼠细胞中表达,所制备DNA疫苗在电子显微镜下可观察到PEG包裹在最外层,粒径均在100 nm以内。成功构建了以PEI磁珠吸附IBRV gB gC gD基因并包裹PEG保护层的DNA疫苗,验证其在小鼠表达系统中可以成功表达,为后续对小鼠免疫效果的评价奠定了基础。To construct an IBRV DNA vaccine by PEI magnetic beads absorbed the tegument protein genes.gB gC gD tegument proteins eukaryotic expression plasmids(pCDNA4-gB-HisB,pCDNA4-gC-His,pCDNA4-gD-His)were amplifier cultured.The PEI magnetic beads were adsorbed and transfected into mouse embryonic cells(NIH/3T3)to identify the protein expression,and then the vaccine was prepared and observed by electron microscopy.The enzyme digest results mean that the eukaryotic plasmids were correct and the WB and IFA.Results showed that the constructed DNA vaccine was proved that the plasmids could be expressed in NIH/3T3 cell line.The DNA vaccine of PEI magnetic beads absorbed IBRV gB gC gD genes was constructed successfully,and could be expressed in NIH/3T3 cell line,which also could be vaccine for mouse.

关 键 词：囊膜蛋白基因 牛传染性鼻气管炎病毒 PEI磁珠 DNA疫苗 PEG600 

分 类 号：S858.23[农业科学—临床兽医学]