单纯疱疹病毒II型gD糖蛋白在真核细胞中的表达

Expression of glycoprotein D of herpes simplex virus type II in eukaryotic cell

目的:构建真核表达质粒pcDNA3/gD,并在体外COS-7细胞中进行表达。方法:体外PCR扩增出gD糖蛋白基因,克隆入真核表达质粒pcDNA3中。PCR、酶切和测序证实插入的gD基因正确后,转染COS-7细胞,用原位ELISA鉴定表达蛋白。结果:构建了pcDNA3/gD真核表达质粒,经原位ELISA证实,转染COS-7细胞表达的gD蛋白能和gD单克隆抗体ID3和DL11结合,证明该蛋白具有天然gD蛋白的抗原性。结论:构建的重组真核表达质粒pcDNA3/gD能够在COS-7细胞中表达。

Objective：To construct recombinant eukaryotic expression vector pcDNA3/gD identified by expression in COS -7 cell. Methods：All code sequence of gD was amplified by PCR and cloned into pcDNA3. After proved to be correct by PCR, enzyme digestion and sequencing, the recombinant vector was transfected into COS- 7 cell, the expressed glycoprotein was detected by in situ ELISA. Results：Recombinant eukaryotic expression vector pcDNAS/gD was constrcted. In situ ELISA result showed that recombinant protein could be identified with gD specific monoclonal antibody. Therefore the protein was of natural antigenic structure of gD. Conclusion：The constructed eukaryotic expression vector pcDNA3/gD could be expressed in COS- 7 cell.