核酶对乙型肝炎病毒基因表达的抑制作用

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ｐＧＥＭ７ｚｆ（一）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ｐＢＢＳ２１２中,重组质粒转染２．２．１５细胞,ＥＬＩＳＡ方法惭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

Ribozyme mediated inhibition of hepatitis B virus gene expression in 2.2.15 cell line

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.