目的　研究核酶抗丙型肝炎病毒(HCV) 的有效切割位点并获得高效、特异、无毒、价廉的HCV 特异性反式核酶。方法　根据文献报道的序列设计、合成HCV5′非结构(NC) 区和核心(C)区的特异性反式核酶基因并分别克隆进入真核细胞表达载体pSV2gpt.CDSRα中,转染HCV 感染的MT2 细胞,用定量逆转录聚合酶链反应(RTPCR) 、核酸杂交等方法测定核酶对HCV 的抑制作用。结果　所用的HCV5′NC 区核酶和C 区核酶对HCV 均有显著的抑制作用,抑制率分别为54 .7 % 和62 .1 % 。两者联合作用时抑制率达78 .8% 。结论　HCV5′NC 区和C 区特异性反式核酶在HCV 感染的体外细胞培养模型中对HCV 复制有明显的抑制作用;双靶位核酶联合使用较单靶位核酶效果更好 Objective To study the efficient cleavage site of ribozyme against hepatitis C virus (HCV) and obtain efficient, specific, non-toxic and inexpensive HCV specific trans ribozyme. Methods According to the sequence reported in the literature, a specific trans ribozyme gene of HCV 5 ’non-structural (NC) region and core (C) region was synthesized and cloned into the eukaryotic expression vector pSV2-gpt.CD-SRα Infected with MT 2 cells infected with HCV, quantitative reverse transcription-polymerase chain reaction (RT PCR), nucleic acid hybridization and other methods to determine the ribozyme inhibition of HCV. Results The HCV 5’nuclear ribozyme and the C-ribozyme used in this study showed significant inhibitory effect on HCV with the inhibition rates of 54.7% and 62.1%, respectively. The combined effect of the two inhibition rate of 78.8%. Conclusion The specific ribo-ribozymes in HCV5’NC region and C region have a significant inhibitory effect on HCV replication in HCV-infected in vitro cell culture model; the combination of dual-target ribozyme and single-target ribozyme is more effective