摘要
将Ｅｐｓｔｅｉｎ－Ｂａｒ（ＥＢ）病毒主要的膜抗原（ＭＡ）ＢＬＬＦ１基因片段插入ｐＨＤ１０１－３质粒的ＣＭＶ启动子下游，构建了真核表达质粒ｐＨＤ－ｇｐ３５０，并转染２９３细胞进行瞬间表达。用免疫荧光法从细胞膜检测到表达的抗原能与其单克隆抗体发生特异性结合，Ｗｅｓｔｅｒｎ－ｂｌｏｔ法证实，表达的抗原分子量为３５０ｋＤ．用能在真核细胞表达的重组质粒ｐＨＤ－ｇｐ３５０的ＤＮＡ，经Ｓｅｐｈａｒｏｓｅ２Ｂ柱纯化后，注射经普鲁卡因预处理的Ｂａｌｂ／Ｃ小鼠的四头肌，观察到ＥＢＶ－ＩｇＡ／ＭＡ抗体水平比ＥＢＶ－ＩｇＧ／ＭＡ低，而ＥＢＶ－ＩｇＡ／ＭＡ的持续时间比ＥＢＶ－ＩｇＧ／ＭＡ长。采用表达ＥＢＶＭＡ的质粒ＤＮＡ与ＣＨＯ细胞表达的ＭＡ蛋白免疫小鼠，均获得抗ＥＢＶＭＡ的抗体。
A plasmid (pHD-gp350) expressing the Epstein-Barr virus membrane antigen (EBV-MA) encoded by the BamHI-L fragment of EBV was constructed.The EBV-MA coding region located on the pVL1393-gp350 plasmid was inserted into the pGEM derivative pHD101-3,where gene expression is driven by the cytomegalovirus(CMV) immediate-early promoter.The MA gene was transiently expressed in 293 cells,and the protein band of gp350kD was found on Western-blot.Detection of MAgp350 was proved by immunofluorescence (IF) test. After purification,this plasmid was directly injected into the quadriceps muscle of Balb/C mice.EBV-IgG/MA and EBV-IgA/MA antibodies could be detected by ELISA.More than 90% of mice had IgG/MA and IgA/MA antibodies,but they disappeared 8-10 week safter injection.Our results showed that injection of purified EBV MAgp350 glycoprotein and injection of pHD-g p350 DNA were both able to elicitanimmune response in mice.
出处
《病毒学报》
CAS
CSCD
北大核心
1997年第1期41-46,共6页
Chinese Journal of Virology
