【摘要】 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更多还原

【Abstract】 Objective To determine the cleavage sites of ribozymes against hepatitis C virus(HCV), and to obtain a highly effective, nontoxic and inexpensive antisense ribozyme specific for HCV. Methods Two effective ribozymes, targeted to HCV 5′ non coding region(5′ NCR) and C region, were synthesized. Eukaryotic expression vectors, pSV2 gpt. CD SRα, containing either HCRZNC or HCRZC were constructed and transfected into MT 2 cells, which had been infected by HCV. Quantitative RT PCR and hybridization methods were used to determine the effect of inhibition of HCV by ribozymes. Results HCRZNC and HCRZC suppressed the replication of HCV by 54.7% and 62.1%, respectively. Furthermore, when the two ribozymes were cotransfected into cells, they suppressed replication by 78.8%. Conclusion Two specific antisense ribozymes have strong inhibitory effects on the replication of HCV in cultured cells, and have better effect when used together.更多还原