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目的：构建针对乙型肝炎病毒（HBV）C区基因（2029-31及2063-65位点）的双位点核酶的真核表达载体．观察其在细胞内对HBV基因表达的抑制作用．方法：利用亚克隆技术．从质粒pGEMRzl23切下EcoRI-BamHI片段，双粘端克隆于真核质粒pBBS212中，将该质粒与P1．2Ⅱ（含HBV全序列）共转染HHCC细胞（人肝癌细胞株）．观察该核酶对HBV基因表达的抑制作用．结果：在HHCC细胞中p1．2Ⅱ可表达出HBsAg及HBeAg该核酶对HBeAg的合成抑制率达65％．结论：该双位点核酶可能通过针对HBVC区mRNA的剪切作用，阻断C区mRNA的表达．抑制了HBeAg的合成。
OBJECTIVE: To construct an eukaryotic expression vector for the double-site ribozyme against hepatitis C virus (HBV) C region (2029-31 and 2063-65). Observe the inhibition of HBV gene expression in the cell. Methods: Using subcloning techniques. The EcoRI-BamHI fragment was excised from the plasmid pGEMRzl23. The double-stick clone was cloned in the eukaryotic plasmid pBBS212. The plasmid was cotransfected into HHCC cells (human hepatoma cell line) with P1.2II (complete sequence containing HBV). To observe the inhibitory effect of this ribozyme on HBV gene expression. Results: The expression of HBsAg and HBeAg in p1.2Ⅱ in HHCC cells could inhibit the synthesis of HBeAg by 65%. Conclusion: The double-site ribozyme may block the expression of C region mRNA by shearing the mRNA of HBVC region. Inhibits the synthesis of HBeAg.