摘要
目的 探讨抗丙型肝炎病毒 (HCV)核酶 (Rz)预先转染的细胞内免疫效应 .方法 应用从前构建的 HCV锤头结构 Rz(Rz2 13) ,通过脂质体介导的基因转染方法 ,转染人肝癌细胞 (HHCC) ,经 G418筛选转染 Rz2 13的基因克隆 ,应用 luc报告基因的表达活性 ,观察该细胞克隆对靶基因 (p CMVNCRluc)转染的抑制作用 .结果 G418筛选能使 Rz2 13在转染细胞内有效表达 Rz RNA,转染 Rz的多克隆细胞能有效地阻断靶基因的再转染 .结论 我们构建的 Rz2 13真核表达载体能在 HHCC细胞内有效表达 ,预先转染发挥细胞内免疫作用 ,可防止 HCV感染 .
AIM To determine the protective efficacy of hammerhead ribozymes (Rz 213) against hepatitis C virus (HCV) from viral gene transfection. METHODS Rz213 cleaving 5 noncoding region (5NCR) of HCV line was beforehand transfected in a human hepatic carcinoma cell (HHCC) and selected for G418 resistance. Cells stably expressing Rz213 were retransfected by pCMVNCRluc containing 5NCR luc fusion genes by lipofectAMINE, and then the luciferase activity in the lysis was measured by scintillation counter. RESULTS HHCC cells stably expressing Rz213 exhibited significant resistance to retransfection of targeting gene. CONCLUSION The ribozymes 213 against the HCV 5NCR could substantially and specifically be expressed in HHCC, which performs an intracellular immunizing function.
出处
《第四军医大学学报》
2000年第7期796-798,共3页
Journal of the Fourth Military Medical University
基金
全军医药卫生基金!资助项目 ( 89D0 48)
1999年第二届国际病毒性肝炎大会报告
关键词
丙型肝炎病毒
锤头结构核酶
细胞内免疫
hepatitis C virus
hammerhead ribozyme
intracellular immunization
