摘要
目的 探讨核酶在细胞内抑制乙型肝炎病毒基因表达的作用 .方法 计算机设计针对 C基因的三个切点的核酶Rz1,Rz2 ,Rz3,合成核酶基因并将三个核酶基因两两组合克隆入 p GEM7zf(- )质粒中 ,经体外转录后切割靶 RNA;选择Rz2和 Rz3串联的双位点核酶基因构建入逆转录病毒载体p BBS2 12中 ,重组质粒转染 2 .2 .15细胞 ,EL ISA方法分析核酶对乙型肝炎病毒基因表达的抑制作用 .结果 三种不同组合的核酶均可有效切割乙型肝炎病毒基因体外转录底物 .2 .2 .15细胞内表达的双位点核酶可明显抑制 HBe Ag的表达 ,抑制率为 48.6 % ,但对 HBs Ag无明显的抑制作用 .结论 核酶可抑制细胞内乙型肝炎病毒基因的表达 .
AIM To investigate inhibition of hepatitis B virus (HBV) gene expression in 2.2.15 cell line by ribozyme. METHODS Three single ribozymes (Rz1, Rz2, Rz3) targeted on core gene of HBV were designed with the computer, then three dual target ribozymes genes were synthesized and cloned in transcript vector pGEM7zf (-). In vitro , cleavage activities of dual target ribozymes were observed with radioautography. To observe inhibition of ribozyme, we cloned Rz23 ribozyme gene into eukaryotic expression vector pBBS212, then transfected it in 2.2.15 cells with lipofectamine. The s and e/c antigens of HBV were detected by ELISA method. RESULTS Three dual target ribozymes could cleave RNA of core gene of HBV respectively; Rz23 ribozyme could be expressed on 2.2.15 cell and inhibit expression of core gene, but there were no effects on expression of s antigen. CONCLUSION Ribozyme can inhibit HBV gene expression in 2.2.15 cells.
出处
《第四军医大学学报》
2000年第7期787-789,共3页
Journal of the Fourth Military Medical University
基金
国家自然科学基金!资助项目 ( 3 95 70 65 2 )
关键词
乙型肝炎病毒
核酶
基因表达
hepatits B virus
ribozyme
gene expression
