表达EB病毒主要膜蛋白gp340/220不同蛋白区段的非复制型重组痘苗病毒的构建  

Construction of Non-replicating Recombinant Vaccinia Virus Expressing Different Protein Fragments of Epstein-Barr Virus Major Membrane Protein gp340/220

作　　者：齐建国[1] 谷淑燕[1] 王卫东[1] 周为民[1] 阮力[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京100052

出　　处：《病毒学报》2004年第1期22-27,共6页Chinese Journal of Virology

基　　金：国家 8 63高科技研究发展计划 ( 863 -10 2 -0 7-0 2 -0 4)

摘　　要：Epstein Barr病毒 (EBV)主要膜蛋白 gp340 / 2 2 0 ,是研究EBV亚单位疫苗的主要抗原。为了更详细地研究gp340 / 2 2 0的免疫原性 ,用PCR法获得不同区段的EBVBLLF1基因 ,并将其克隆入表达载体 pNeock1 1 β7 5中 ,通过重组质粒与非复制痘苗病毒在鸡胚成纤维细胞中同源重组 ,获得 3株表达EBVgp340 / 2 2 0不同区段的重组痘苗病毒。PCR和Southernblot结果证实 ,各个重组病毒基因组中有相应外源基因的整合。Westernblot结果表明 ,3种重组病毒感染人源细胞后都能够稳定表达蛋白 ,表达蛋白均为糖蛋白。免疫荧光结果表明 ,3种蛋白分布在细胞的不同部位。重组痘苗病毒的成功构建为更好地研究gp340 / 2 2The Epstein Barr virus(EBV)major membrane protein,gp340/220,was known to be a candidate subunit vaccine for EBV.To further investigate the immunogenicity of gp340/220,different DNA fragments of EBV BLLF1 gene were amplified by PCR technology and cloned into the expression vector pNeock11β7 5.Three non replicating recombinant vaccine viruses expressing different protein fragments of gp340/220 were constructed through the homologous recombination between recombinant plasmids and non replicating vaccinia virus in chicken embryo fibroblast(CEF)cell.PCR and Southern blot analysis revealed that each foreign gene was integrated into its relative recombinant vaccinia virus genome.Western blot demonstrated that the corresponding proteins were stably expressed in cells infected with different recombinant viruses.All products are glycoproteins.Immunofluorescence analysis showed that each protein expressed by recombinant virus was localized in different parts of the cell.These results provide detailed experimental information to elucidate the immunogenicity of gp340/220.

关 键 词：EPSTEIN-BARR病毒 EBV 疱疹病毒科 膜蛋白 非复制型重组痘苗病毒 

分 类 号：R373[医药卫生—病原生物学] Q782[医药卫生—基础医学]