MS2介导的口蹄疫类病毒颗粒疫苗的研究  

Study on MS2 Mediated Virus-like Particles Vaccine Against Foot-and-month Disease

作　　者：董艳美[1] 张国广[1,2] 汪卫[1] 范红军[3] 陈亮[1] 

机构地区：[1]厦门大学生命科学学院,福建厦门361102 [2]漳州师范学院生命科学与技术系,福建漳州363000 [3]河南师范大学生命科学学院,河南新乡453000

出　　处：《厦门大学学报（自然科学版）》2013年第2期237-243,共7页Journal of Xiamen University：Natural Science

基　　金：福建省科技厅重点项目(2009N0051);福建省自然科学基金项目(2010J01240);厦门市科技计划项目(3502Z20103007)

摘　　要：研究表明,口蹄疫病毒(Foot-and-mouth disease virus,FMDV)的VP1蛋白含有关键的抗原决定簇.根据O/HL-JOC12/03猪源FMDV毒株VP1上第141～160位氨基酸序列设计引物,利用重叠延伸PCR(SOE PCR)技术将该序列插入在大肠杆菌(Escherichia coli)噬菌体MS2外壳蛋白基因的特定位点.然后连接到原核表达载体pET28a上,构建了重组表达质粒28a-CP-VP1,转化BL21(DE3),ITPG诱导表达得到融合表达的CP-VP1.透射电子显微镜下观察融合蛋白CP-VP1成类病毒颗粒(virus-like particles,VLPs)状.间接ELISA实验证实该蛋白能够特异地结合豚鼠抗O型FM-DV的阳性血清,30μg的CP-VP1的VLPs蛋白免疫小鼠后可以诱导其产生高效价的特异抗体.故该口蹄疫病毒抗原决定簇展示在噬菌体MS2上表面的VLPs蛋白具有开发成口蹄疫疫苗的前景.Foot-and-mouth disease （FMD） has caused severe economic losses to millions of farmers in many countries all over the world.The severity of the disease and the spread of this epidemic virus have not been controlled fruitfully,and it continues to be a threat to livestock.Vaccination is one of the most effective weapons against the infectious disease.Our study was aimed to develop a kind of effective and safe virus-like particles （VLPs） vaccine.VP1 protein of foot-and-mouth disease virus （FMDV） has key antigenic determinants which were used by vaccine developers time and again.The nucleotides coding the peptides from 141 to 160 AA of VP1 protein of strain HLJOC12/03 FMDV were amplified and inserted into the specific site in the coat protein gene of MS2 by using SOE PCR.The products were digested by enzymes and then ligated to the pET28a.The recombination plasmids were transformed into Escherichia coli BL21 （DE3） host cell,expressed with IPTG induction,and the high level expression of the protein were harvested.The recombinant CP-VP1 virus-like particles （VLPs） protein was observed by transmission electron microscopy.A series of immunogenicity tests were carried out and the experimental data proved that the VLPs protein we had designed showed fine effects and it would be a kind of hopeful vaccine against FMD,and this kind of VLPs can also be applied to detect FMD.

关 键 词：口蹄疫 MS2噬菌体 抗原决定簇 类病毒颗粒 

分 类 号：Q78[生物学—分子生物学]