疫苗接种是控制禽流感发生的主要措施之一,利用病毒保守的M2蛋白开发具有交叉免疫保护力的基因工程亚单位疫苗。将M2蛋白的膜外区(M2E表位)与HbCAg采用重叠延伸PCr技术构建融合基因M2EHbC+,然后将融合基因进行原核表达并检测了表达产物的动物免疫效果。结果表明,融合基因M2EHbC+能在原核系统中高效表达,表达蛋白能够被M2抗体识别,具有良好的抗原性,纯化出的重组蛋白可以自主包装成病毒样颗粒结构,能诱导小鼠产生M2E特异性抗体。M2E表位在重组蛋白颗粒中得到有效展示,为开发具有交叉保护力的禽流感亚单位疫苗打下了基础。The present vaccination is one of the main strategies for the control of avian influenza virus(AIV).Previous investigations had shown that subunit component vaccine based on M2 gene or M2e epitope could fully protect mice against a lethal H5N1 subtype avian influenza virus infection,because the extracellular domain of the minor virus-coded M2 protein is nearly invariant in all influenza A strains.Numerous studies have been devoted to developing novel heterosubtypic cross-protection vaccine by using M2 gene or M2e epitope.In this study,the M2e domain to N terminal and major immunodominant region of the hepatitis B virus core(HBcAg) protein was genetically fused to create fusion gene coding M2eHBc+ by overlap extension PCR.M2eHBc+ was expressed in Escherichia coli cell afterwards and the immune efficacy was detected in mice with pure expressing products.Fusion gene M2eHBc+ could be efficiently expressed in E.coli cell,and the expressed peptide was in soluble form and possessed M2 antigenicity since peptide can be detected by M2 antibody.The purified M2eHBc+ protein using affinity chromatography can self-assemble into virus-like particles(VLP) and can induce anti-M2e IgG in the serum of immunized mice.M2e epitope was effectively displayed on the outer surface of VLP,which laid the foundation for further development of recombinant subunit AIV vaccine.福建省科技厅重大项目子课题(2006NZ0003-2);福建省科技厅重点项目(2009N0051
