乙肝病毒X基因可调控表达细胞模型的构建  

Cell Model Construction of Regulated Hepatitis B Virus X Gene Expression

作　　者：白荷露[1] 刘蕊[1] 朱乃硕[1] 

机构地区：[1]复旦大学生命科学学院微生物学与微生物工程系,上海200433

出　　处：《生物技术通报》2013年第2期140-145,共6页Biotechnology Bulletin

基　　金："艾滋病和病毒性肝炎等重大传染病防治"科技重大专项(2008ZX10002-002);国家自然科学基金项目(30901315)

摘　　要：利用四环素诱导型慢病毒系统(Lentil-XTMTet-On Advanced Inducible Expression System)构建了含有乙肝病毒X基因(HBx)的可诱导表达载体质粒,利用慢病毒包装细胞系HEK293T细胞分别获得含有调控元件rt-TA和HBx基因的病毒颗粒,滴度达到108LPs/mL。将两种病毒先后感染HepG2肝癌细胞系,分别利用G418和Puromycin抗生素筛选获得整合有HBx基因的稳定细胞株。在体系中加入Dox时,应用RT-PCR和Western blot方法验证了HBx基因的可诱导表达。可诱导表达细胞模型构建成功,为进一步研究HBx基因在对宿主感染致病和免疫耐受中的作用提供了一种较理想的试验模型。An HBx inducible expression plasmid carrier was constructed by Lentil-XTM Tet-On Advanced Inducible Expression System, and rt-TA gene virus particles and HBx gene virus particles were synthesized by lentiviral packing cell line HEK293T cells, with the titer reaching 10^8 LPs/mL. When HepG2 cell lines were infected by the two virus particles and selected by G418 and puromycin antibiotics, stable cell line with HBx gene could be constructed. After adding Dox into the stable cell line, the inducible expression of HBx gene could be proved by RT-PCR and Western blot. Our study provides an ideal experiment model for further research on the roles of HBx in HBV infection and the host immune tolerance.

关 键 词：乙型肝炎病毒基因 慢病毒表达载体 四环素调控 可诱导表达 

分 类 号：R373.21[医药卫生—病原生物学]