HCV E2和HBV preS融合蛋白真核表达载体的构建及在哺乳动物细胞中的表达

Constructing the eukaryotic expression vector expressing HCV E2 protein combined with HBV preS protein and expressing the combined protein in mammalian cells

目的：构建丙型肝炎病毒（ＨＣＶ）Ｅ２和乙型肝炎病毒（ＨＢＶ）ｐｒｅＳ融合蛋白的真核表达载体。方法：用ＰＣＲ方法扩增ＨＢＶｐｒｅＳ和ＨＣＶＥ２蛋白基因，并将二者克隆到真核表达载体ｐｃＤＮＡ３中，用脂质体转染试剂将其转入ＣＯＳ７细胞，通过免疫荧光检测细胞瞬时表达的融合蛋白。结果：Ｅ２ｐｒｅＳ融合蛋白基因包括了全长的ＨＢＶｐｒｅＳ和ＨＣＶＥ２蛋白基因，嵌合基因长度约１．６ｋｂ。表达载体在ＣＯＳ７细胞表达出了Ｅ２ｐｒｅＳ融合蛋白。结论：构建的Ｅ２ｐｒｅＳ融合蛋白表达载体能在哺乳动物细胞中表达融合蛋白。

Objective: To construct the eukaryotic expression vector which expresses the combined protein of HCV E2 and HBV preS proteins. Methods: HCV E2 and HBV preS genes were amplified with PCR and cloned into mammalian expression vector pcDNA3. The constructed vector was transfected into COS7 cells with lipofectin. The expressed E2 preS protein was detected by means of immunofluorescence. Results: The chimeric gene, which was about 1.6 kb, included entire HBV preS and HCV E2 gene. The cells transfected with the constructed vector expressed E2 preS protein successfully. Conclusion: The constructed vector containing chimeric gene of E2 preS protein can express E2 preS protein in mammalian cell. The success of constructing this vector laid the foundation for obtaining the combined protein and studying its immunogenicity.