乙型肝炎病毒蛋白和绿色荧光蛋白的共表达研究  

Co-Expression of Hepatitis B Virus Proteins and Green Fluorescent Protein After Eukaryotic Cell in vitro and Hydrodynamics-Based in vivo Transfection

作　　者：黄洁[1] 訾小渊[1] 韩庆旺[1] 李建秀[1] 王新民[1] 杨勇骥[2] 胡以平[1] 

机构地区：[1]第二军医大学细胞生物学教研室,上海200433 [2]第二军医大学生物物理研究所,上海200433

出　　处：《生物技术通讯》2008年第2期167-170,共4页Letters in Biotechnology

摘　　要：目的:构建增强型绿色荧光蛋白(EGFP)标记的乙型肝炎病毒(HBV)真核表达载体,并研究其在真核细胞和小鼠体内的共表达。方法:以质粒pBR322-HBVadr2.0和pCX-EGFP为基础,构建含有双拷贝HBV全基因组DNA和EGFP基因的真核表达载体pCX-EGFP-HBVadr2.0,分别转染真核细胞和小鼠肝组织,建立体外、体内表达系统,研究GFP和HBV基因的表达。结果:构建了真核表达载体pCX-EGFP-HBVadr2.0,EGFP和HBV病毒蛋白在体内和体外均可表达。结论:构建的pCX-EGFP-HBVadr2.0真核表达载体可以GFP作为HBV存在与否的报告基因,提高了培育检测转基因小鼠的效率,为转基因小鼠的制备及后续研究奠定了基础。Objective： To construct pCX-EGFP-HBVadr2.0 eukaryotic expression vector harboring 2 copies of hepatitis B virus（HBV） genomic DNA and enhanced green fluorescent protein（EGFP） gene and investgate the co-expression in vitro and in vivo. Methods： The eukaryotic expression vector pCX-EGFP-HBVadr2.0 was constructed based on plasmids of pBR322-HBVadr2.0 harboring 2 copies of HBV genomic DNA and vector pCX-EGFP containing CMV-IE enhancer, chicken B-action promoter and EGFP by recombination DNA technique. The HBV genomic DNA and EGFP gene expressing in transfected L02 cell line in vitro and hydrodynamic injected mouse in vivo were studied. Results： The eukaryotic expression vector pCX-EGFP-HBVadr2.0 was constructed, and could expressed in vitro and in vivo. Conclusion： The HBV eukaryotic expression vector carrying EGFP reporter gene to generate transgenic mice was constructed. It should be practicable and effective in breeding and estimation by using HBV transgenic mice with the EGFP.

关 键 词：乙型肝炎病毒 增强型绿色荧光蛋白 真核表达载体 转基因小鼠 

分 类 号：Q78[生物学—分子生物学]