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目的选择针对丙型肝炎病毒（HCV）5’非编码区（NCR）和核心区（C）的核酶（ribozyme，Rz）切割位点，构建带自剪切的Rz真核表达载体。方法应用计算机辅助设计，根据能量最低化原理，以HCV－H（1a）株SNCR和C靶序列，预测其二级结构，选择理想的Rz切割位点，设计锤头结构核酶；体外合成针对HCV5’NCR的Rz213和Rz260的cDNA，通过亚克隆技术连接于真核表达载体（pcDNA3）。结果在124个自然切割位点（CUX和GUX）中选出213（CUC）、260（GUA）、407（GUC）和498（CUU）4个切点；Rz213和Rz260的DNA序列分析，结果与合成序列完全一致，酶切鉴定两Rz连接正确。结论计算机可作为抗病毒Rz设计的重要辅助工具；通过亚克隆技术可使目的Rz两端带自剪切Rz，并成功地插入真核表达载体。
Objective To select ribozyme (Rz) cleavage site for 5 ’non-coding region (NCR) and core region (C) of Hepatitis C virus (HCV) to construct self-sheared Rz eukaryotic expression vector. Methods The computer-aided design was used to predict the secondary structure of HCV-H (1a) SNCR and C target sequences according to the principle of energy minimization. The ideal Rz cleavage site was selected to design a hammerhead ribozyme. CDNA of Rz213 and Rz260 of HCV 5 ’NCR was ligated to eukaryotic expression vector (pcDNA3) by subcloning technique. Results Four pointcuts of 213 (CUC), 260 (GUA), 407 (GUC) and 498 (CUU) were selected from 124 natural cleavage sites (CUX and GUX). DNA sequence analysis of Rz213 and Rz260 showed that The synthetic sequences were identical, and the two Rz connections were identified by restriction enzyme digestion. Conclusion The computer can be used as an important auxiliary tool for antiviral Rz design. By subcloning technology, Rz can be self-sheared both ends of Rz and successfully inserted into eukaryotic expression vector.