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目的探讨C基因截短突变体抗乙型肝炎病毒(HBV)的作用机制。 方法 构建C基因截短的真核表达载体pcDNA3—△ C及野生型C基因真核表达载体pcDNA3—C,瞬时转染HepG2细胞,用SDS—PAGE western blot检测pcDNA3—△C、pcDNA3-C的蛋白表达。pcDNA3—△ C与adwR9共转染HepG2细胞,以pcDNA3与adwR9为对照,用荧光定量PCR检测培养上清液及细胞内病毒量,用Native western blot 分析C基因截短蛋白干扰核心颗粒形成。 结果 重组载体pcDNA3—△ C、pcDNA3—C均可表达,pcDNA3-△C与adwR9共转染组上清液和细胞内病毒量较对照组降低,pcDNA3—△ C和pcDNA3—C共转染组Native western blot条带与pcDNA3和pcDNA3—C共转染组条带相比较明显淡。 结论 C基因截短突变体可干扰核心颗粒的形成,导致HBV复制下降。
Objective To investigate the mechanism of anti-hepatitis B virus (HBV) truncated mutant of C gene. Methods The truncated eukaryotic expression vector pcDNA3- △ C and wild type C gene eukaryotic expression vector pcDNA3-C were constructed and transfected into HepG2 cells transiently. The expression of pcDNA3- △ C, pcDNA3-C Protein. The pcDNA3- △ C and adwR9 co-transfected HepG2 cells, pcDNA3 and adwR9 as control, the fluorescence quantitative PCR detection of the supernatant and intracellular viral load, using Native western blot analysis of C gene truncated protein interferes with the formation of core particles. Results The recombinant plasmids pcDNA3- △ C and pcDNA3-C were both expressed in the supernatant and the cells in the co-transfection group of pcDNA3- △ C and adwR9 compared with the control group. The co-transfection of pcDNA3- △ C and pcDNA3-C Native western blot bands were significantly weaker than those of pcDNA3 and pcDNA3-C co-transfection groups. Conclusion C truncated mutants can interfere with the formation of core particles, leading to a decrease in HBV replication.