插入CpG序列和IL-2基因的HBV重组真核表达载体的构建  

CONSTRUCTION OF RECOMBINANT HBV MAMMALIAN EXPRESSION VECTORS BY INSERTION OF CpG MOTIF AND IL-2 GENE RESPECTIVELY

作　　者：罗红雨[1] 杨旭[1] 苏先狮[1] 

机构地区：[1]中南大学湘雅二医院传染科,长沙410011

出　　处：《中国现代医学杂志》2001年第7期25-27,30,共4页China Journal of Modern Medicine

基　　金：湖南省自然科学基金重点项目资助 (98JJY10 0 3 )

摘　　要：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 。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.

关 键 词：乙型肝炎 基因 疫苗 重组 质粒 CPG序列 IL-2基因 HBV 

分 类 号：R512.6[医药卫生—内科学]