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Inhibition of HBV replication by M1GS RNA ribozymes targeting the S and C regions of the HBV genome
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- Author:
- No author available
- Journal Title:
- WORLD CHINESE JOURNAL OF DIGESTOLOGY
- Issue:
- 28
- DOI:
- 10.3969/j.issn.1009-3079.2007.28.006
- Key Word:
- 乙型肝炎病毒;核酶;核糖核酸酶P;基因治疗;逆转录聚合酶链式反应;酶联免疫吸附试验
Abstract： 目的:研究靶向HBV S区和C区基因的M1GSRNA核酶共同作用对HBV基因表达的影响.方法:选择HBV ayw亚型S区基因294 nt和C区基因2333 nt为切割位点,以含有编码M1 RNA的DNA序列的质粒pTK117为模板.通过PCR扩增得到M1GS RNA核酶的DNA模板,并将其克隆至真核表达载体pEGFP-C1得到重组质粒pEGFP-GSS和pEGFP-GSC.将2个重组质粒共转染HepG2.2.15细胞,转染后ELISA法测细胞培养液中的HBsAg和HBeAg,RT-PCR检测HBV mRNA.结果:成功构建了分别靶向HBV S区基因和C区基因的真核表达载体.共转染HepG2.2.15细胞后,HBsAg和HBeAg的表达分别被抑制了33.2%和39.1%.HBV C mRNA和S mRNA分别被抑制了32.5%和29.7%,而HepG2.2.15细胞的增殖无明显变化.结论:靶向HBV S区和C区基因的M1GS RNA核酶共同作用可特异性抑制HBV S区和C区基因的表达.
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