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[肝富集转录因子结合位点突变在乙型肝炎病毒基因组C启动子区域对HBx增强乙型肝炎病毒复制中的作用]

[Role of the liver-enriched transcription factor binding site mutation in the C promoter region of hepatitis B virus genome for HBx-enhanced hepatitis B virus replication].

作者信息

Han N, Yan L L, Du L Y, Huang F J, Tang H

机构信息

Center of Infectious Diseases, West China Hospital of Sichuan University, Chengdu 610041, China.

West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu 610041, China.

出版信息

Zhonghua Gan Zang Bing Za Zhi. 2021 Apr 20;29(4):350-355. doi: 10.3760/cma.j.cn501113-20200923-00523.

Abstract

To construct a recombinant HBV replication-type plasmid with liver-enriched transcription factor binding site mutation at proximal of HBV C promoter in order to elucidate the role of HBx-enhanced HBV replication. Site-directed mutagenesis technology was used to construct a recombinant plasmid with liver-enriched transcription factor binding site mutation at proximal of HBV C promoter on the basis of wild-type HBV replicating plasmid and HBV replicating plasmid lacking HBx expression. Subsequently, plasmid transfection was carried out in HBV liver cancer cell replication model and mouse replication model, and HBV replication intermediates of cells and mouse liver tissue were extracted for detection. Based on the HBV replicating plasmid, the HBV replicating plasmid with liver-enriched transcription factor binding site mutation at proximal of HBV C promoter was successfully constructed. HBx-enhanced HBV replication were detected in both the HBV liver cancer replication model and the mouse replication model. After mutating liver-enriched transcription factor binding site mutation at proximal of HBV C promoter, the effect of HBx on the enhancement of HBV replication was not significantly affected. HBx may not enhance HBV replication through liver-enriched transcription factor binding site mutation at proximal of HBV C promoter. The role of other liver-enriched transcription factor binding sites in HBx-enhanced HBV replication needs further study.

摘要

构建一种在HBV C启动子近端具有肝富集转录因子结合位点突变的重组HBV复制型质粒,以阐明HBx增强HBV复制的作用。利用定点诱变技术,在野生型HBV复制质粒和缺乏HBx表达的HBV复制质粒的基础上,构建一种在HBV C启动子近端具有肝富集转录因子结合位点突变的重组质粒。随后,在HBV肝癌细胞复制模型和小鼠复制模型中进行质粒转染,并提取细胞和小鼠肝组织的HBV复制中间体进行检测。基于HBV复制质粒,成功构建了在HBV C启动子近端具有肝富集转录因子结合位点突变的HBV复制质粒。在HBV肝癌复制模型和小鼠复制模型中均检测到HBx增强的HBV复制。在HBV C启动子近端的肝富集转录因子结合位点发生突变后,HBx对HBV复制增强的作用未受到明显影响。HBx可能不是通过HBV C启动子近端的肝富集转录因子结合位点突变来增强HBV复制的。其他肝富集转录因子结合位点在HBx增强HBV复制中的作用有待进一步研究。

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