摘要
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 explore the effect and mechanism on HBV replication in C gene truncated mutant. Methods Protein expression of C gene truncated vector and wild C gene vector were assay by SDS-PAGE Western blot. Constructed C gene truncated expression vector was cotransfected with wild HBV genome; virus load was detected by PCR in the culture medium and the cell. The formation of core particle was assay by Native western blot. Results The recombinant vectors can efficiently express. Virus load of the cotransfected group by pcDNA3- AC and adwR9 was lower than that of control group in the culture medium and the cell. Protein band of the co-expressed group by pcDNA3- AC and pcDNA3-C showed slightly weaker than that of the co-expressed group by pcDNA3 and pcDNA3-C. Conclusion C gene truncated mutant could interfere with the formation of core particle and reduce of HBV replication.
出处
《中华肝脏病杂志》
CAS
CSCD
2004年第5期290-292,共3页
Chinese Journal of Hepatology
基金
国家自然科学基金(30371287)
