插入CpG序列和IL—2基因的HBV重组真核表达载体的构建

目的：构建插入CpG序列和IL-2基因的HBV重组真核表达载体。方法：采用PCR产物直接克隆方法，构建了编码HBsAg基因的真核细胞表达载体pcDNA3.1^ -HBsAg；并以此为基础通过重组DNA技术分别构建了在不同位置含不同数目的CpG序列的重组质粒pcDNA3.1^ -S/CpG1、pcDNA3.1^ -S/CpG2、pcDNA3.1^ -S/CpG1 CpG2及IL-2和HBsAg融合基因的表达载体pcDNA3.1^ -S/IL-2。通过脂质体基因转移技术导入Cos-7细胞中检测其瞬间表达。结果：通过酶切、PCR及测序证实已分别正确完整地插入IL-2基因和CpG序列，体外转染Cos-7细胞后可见基因的表达和分泌，结论：本实验成功构建了我们所设计的5种重组质粒，拟为今后探索优化基因疫苗的设计及HBV治疗性疫苗的研究奠定基础。

Construction of the Recombinant HBV Mammalian Expression Vectors by Insertion of CpG Motif and IL-2 Gene Respectively

Objective:To construct the recombinant HBV mammalian expression vectors by insertion of CpG motif and IL-2 gene respectively.Methods:Five recombinant plasmid vectors pcDNA3.1 -S,pcDNA3.1 -S/GpG1,pcDNA3.1 -S/CpG2,pcDNA3.1 -S/CpG1 CpG2,pcDNA3.1 -S/IL-2 was constructed by PCR method and DNA recombinant technique.Then,these recombinant vectors were transfered into Cos-7 cells by cation lyposome and were detected their transient expressing product in vitro.Results:It was confirmed that these recombinant plasmid vectors were constructed successfully by the method of restriction endonucleases digestion,PCR and DNA sequencing.Their transient expressing products were detected in virto.Conclusions:Our test may provide basis for further study of optimization HBV DNA vaccine.