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.
出处
《生物技术通讯》
CAS
2008年第2期167-170,共4页
Letters in Biotechnology
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
hepatitis B virus
enhanced green fluorescent protein
transgenic mice
eukaryotic expression vector
