乙型肝炎病毒前S2/S与重组人GM-CSF融合基因的分子克隆及鉴定

目的为探讨乙型肝炎病毒前S2(preS2)和粒细胞-巨噬细胞集落刺激因子(GM—CSF)对乙型肝炎疫苗的免疫增强作用，构建S2／S(preS2 HBsAg)与GM—CSF融合基因真核表达载体。方法采用PCR技术分别扩增出S2／S大小约846bp的编码基因片段和带有15个甘氨酸接头序列的GM—CSF约450bp编码基因片段，经T-A克隆后分布克隆至载体PcDNA3．1中，获得PcDNA3．1-S2／S-GM-CSF融合基因表达载体，利用酶切、PCR和DNA序列测定进行鉴定插入片段的大小和方向。结果经过鉴定该融合基因全长约1300bp，证实为S2／S-GM-CSF融合基因。结论乙型肝炎病毒(HBV)S2／S和GM—CSF融合基因真核表达载体构建成功，为进一步研究其表达和生物学活性奠定了一定基础。

Cloning and identification of fusion gene encoding HBV S2/S and recombinant human GM-CSF

Objective The eukaryotic coexpression plasmid encoding HBV S2/S and recombinant human GM-CSF was constructed to study the immune enhancement of GM-CSF and PreS2 as hepatitis B vaccine.Methods HBV S2/S and recombinant human GM-CSF fragments was amplified by PCR respectively.Then the PCR products were purified and cloned into PcDNA3.1 after T-A cloning.The recombinant plasmid PcDNA3.1-S2/S-GM-CSF identified by restriction enzyme analysis,PCR amplification and/or DNA sequencing.Results The fusion gene determined is S2/S-GM-CSF sequencing of 1300bp.Conclusion The successful construction of S2/S-GM-CSF fusion gene cloned into eukaryotic coexpression plasmid provides the basis for further study of its expression and biological activity.