双位点核酶细胞内抑制乙型肝炎病毒基因的表达

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基因表达抑制作用。方法：利用亚克隆从质粒ｐＧＥＭＲｚ１２３切下ＥｃｏＲ－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Ｐ１．２

Inhibition of dual-target ribozyme against HBV on HBeAg in HHCC cells

AIM: To construct a eukaryotic expression vectorwith dual--target ribozyme against hepatitis B virlls and to observe the inhibition of HBeAg expressed in the HHCC cell(human hepatic carcinoma cell line ) transfected by HIVgenome and dual--target ribozyme against HBV. METHODS:Using subclone technique, the dual--target ribozyme gene cutfrom pGEM-Rz 123 was ligated into the eukaryotic expression vector pBBSZ12. The recombinant plasmid pBBS212uRzand pl. 2 1 carrying genome of adr--subtype HllV were cotransfected into the HHCC cell by using lipofectamine. Theintracellular e/c antigen of HBV was detected by EI.ISA andimmunohistochemistry. RESULTS: The cotransfected cellsdid express HBV e antigen and ribozyrne RNA, and the dualtarget ribozyme inhibited the level of HlleAg expressed on cotransfected cells by up to 65%. CONCLUSION: The dualtarget ribozyme can cleave the HBV core region mRNA andinhibit HBV the expression of HBV core region gene.