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目的:探讨多位点自剪切核酶及突变核酶对细胞内HBVmRNA的切割作用.方法:构建5个不同的多位点核酶及突变核酶的真核表达载体,将他们分别与乙型肝炎病毒全基因序列共转染HepG2细胞,用ELISA,共聚焦定量及图像分析的方法观察多位点核酶在细胞内对HBVmRNA切割作用.结果:构建的真核表达载体在细胞内确可表达出多位点核酶,核酶及突变核酶在细胞内对HBV基因的表达均有抑制作用,不同表达载体的抑制率不同,以tRNA启动子的表达载体抑制效率最高,达81%,突变核酶亦有部分反义RNA的抑制效果.结论:抗乙型肝炎病毒核酶在细胞内可抑制HBV基因的表达,不同表达载体其核酶的表达效率不同.
Objective: To investigate the cleavage of intracellular HBV mRNA by multi-site self-cleaving ribozymes and mutant ribozymes.Methods: To construct eukaryotic expression vectors of five different multi-site ribozymes and ribozymes, HepG2 cells were co-transfected with the full-length Hepatitis virus genome and the effect of multi-site ribozyme on HBV mRNA cleavage was observed by ELISA, confocal quantification and image analysis.Results: The constructed eukaryotic expression vector was indeed The expression of multi-site ribozymes, ribozymes and mutant ribozymes inhibited the expression of HBV gene in the cells, and the inhibition rate of different expression vectors was different. The tRNA promoter expression vector had the highest inhibitory efficiency of 81% Mutation ribozymes also showed partial antisense RNA inhibitory effect.Conclusion: The anti-HBV ribozyme can inhibit the expression of HBV gene in cells, and the expression efficiency of different ribozymes in different expression vectors is different.