水痘-带状疱疹病毒糖蛋白E真核表达载体的构建及表达

Construction and Expression of the Eukaryotic Expression Vector Containing Gene of Varicella-Zoster Virus Glycoprotein E

目的 构建水痘 带状疱疹病毒 (VZV )糖蛋白E的真核表达载体并使其在COS7细胞中表达。方法 用PCR方法扩增VZV糖蛋白E基因 ,并将其克隆到真核表达载体 pcDNA 3 .1。用脂质体转染试剂将重组载体转入COS7细胞 ,通过免疫组化法检测转染细胞瞬时表达的重组蛋白。结果 扩增的目的基因包括了全段糖蛋白E基因 ,长度约1.9kb。重组表达载体在COS7细胞中表达出了糖蛋白E。结论 VZV糖蛋白E真核表达载体的成功构建及在COS7细胞中重组蛋白的表达 ,为研究此蛋白的免疫原性和VZV核酸疫苗打下了基础。

Objective To construct the eukaryotic expression vector which express the protein of VZV glycoprotein E in COS7 cells.Methods VZV glycoprotein E genes were amplified with PCR and cloned into the eukaryotic expression vector pcDNA3.1.The constructed vector was transfected into COS7 cells with lipofection.The expressed VZV glycoprotein E was detected by means of immunohistochemistry.Results The amplified gene,which was about 1.9kb, included entire gene of VZV glycoprotein E.The cells transfected with the constructed vector expressed VZV glycoprotein E successfully.Conclusion The constructed vector containing gene of VZV glycoprotein E can express glycoprotein E in COS7 cell.The success of constructing this vector laid the foundation for studying immunogenicity of glycoprotein E and VZV DNA vaccine.