腺相关病毒主要调控蛋白Rep78的表达及其与HBV-X基因启动子的体外结合

首都医科大学学报 ›› 2006, Vol. 27 ›› Issue (4): 423-426.

腺相关病毒主要调控蛋白Rep78的表达及其与HBV-X基因启动子的体外结合

刘天会, 丛敏, 王萍, 唐淑珍, 王宝恩, 贾继东, 尤红

首都医科大学附属北京友谊医院肝病中心

收稿日期:2006-06-16 修回日期:1900-01-01 出版日期:2006-08-24 发布日期:2006-08-24
通讯作者: 尤红

Expression of the Adeno-associated Virus Rep78 as a Major Regulatory Protein and Its Binding with Hepatitis B Virus X Gene Promoter in Vitro

Liu Tianhui, Cong Min, Wang Ping, Tang Shuzhen, Wang Baoen, Jia Jidong, You Hong

Liver Research Center, Beijing Friendship Hospital, Capital University of Medical Sciences

Received:2006-06-16 Revised:1900-01-01 Online:2006-08-24 Published:2006-08-24

摘要/Abstract

摘要： 目的对腺相关病毒(adeno-associated virus, AAV)Rep78蛋白对乙肝病毒X启动子(HBV-X promoter)的可能作用进行初步研究.方法利用pMAL-Rep78(含有Rep78全长基因的表达质粒)在大肠杆菌中表达并纯化得到带有AAVRep78蛋白的MBP-rep78融合蛋白;通过SDS-电泳及免疫印迹(Western blot)鉴定蛋白表达.合成地高辛标记的X启动子3段探针;以非放射性凝胶电泳阻滞实验(EMSA)检测Rep78与HBV-X启动子的结合.结果得到纯化的MBP-rep78融合蛋白;EMSA结果显示Rep78蛋白在体外能够与HBV-X启动子DNA结合,并有剂量依赖关系.结论 AAVRep78可以与HBV-X启动子结合.

关键词: 腺相关病毒, 乙肝病毒, Rep78, 凝胶电脉阻滞实验

Abstract: Objective This study is to investigate the possible effect of adeno-associated virus(AAV) Rep78 on hepatitis B virus X gene(HBV-X).Methods Rep78 protein was produced as a fusion protein with MBP using pMAL-Rep78 plasmid(with full gene of Rep78) in DH-5α.SDS-PAGE and western blot were used to confirm the expression of MBP-Rep78.Electrophoretic mobility shift assay(EMSA) was utilized to detect the binding of MBP-Rep78 protein with HBV-X promoter.Results Purified MBP-Rep78 fusion protein was obtained successfully.EMSA showed Rep78 protein could bind with HBV-X promoter strongly in a dose-dependent manner.Conclusion AAV Rep78 could bind with HBV-X promoter.

Key words: adeno-associated virus, hepatitis B virus, Rep78, electrophoretic mobility shift assay

中图分类号:

R512.6

刘天会;丛敏;王萍;唐淑珍;王宝恩;贾继东;尤红. 腺相关病毒主要调控蛋白Rep78的表达及其与HBV-X基因启动子的体外结合[J]. 首都医科大学学报, 2006, 27(4): 423-426.

Liu Tianhui;Cong Min;Wang Ping;Tang Shuzhen;Wang Baoen;Jia Jidong;You Hong. Expression of the Adeno-associated Virus Rep78 as a Major Regulatory Protein and Its Binding with Hepatitis B Virus X Gene Promoter in Vitro[J]. Journal of Capital Medical University, 2006, 27(4): 423-426.

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