【摘要】 研究表达的丙型肝炎病毒(HCV)NS4B对Hep3B细胞非折叠蛋白质反应的影响。NS4B重组真核表达质粒pcDNA3.1(-)NS4B通过脂质体转染Hep3B细胞,G418筛选和Western blot鉴定稳定转染细胞;RT-PCR检测稳定转染细胞内XBP1 mRNA剪接,Western blot鉴定ATF6蛋白切割,荧光素酶试验检测稳定转染细胞内GRP78和XBP1启动子活性。G418筛选和Western Blot鉴定证实获得稳定表达NS4B的Hep3B细胞;在该细胞内,XBP1 mRNA剪接、ATF6切割、XBP1和Grp78启动子激活均被检测到。NS4B在Hep3B的稳定表达诱导了非折叠蛋白质反应。更多还原

【Abstract】 To study the effect of hepatitis C virus NS4B on unfolded protein response in Hep3B cells.The recombinant vector pcDNA3.1(-) NS4B with NS4B gene was transfected to Hep3B cells using Lipofectamine 2000.Stably transfected cells were selected by G418 and then confirmed by Western blot analysis;XBP1 mRNA splicing,ATF6 cleavage,GRP78 and XBP1 promoter activation were analyzed in stably transfected Hep3B cells using RT-PCR,Western blot or luciferase assays.G418 seclection and Western blot demonstrated that Hep3B cells stably expressing NS4B were obtained successfully;XBP1 mRNA splicing,ATF6 cleavage,GRP78 and XBP1 promoter activation were observed in Hep3B cells stably expressing NS4B.Stable expression of NS4B in Hep3B cells induces unfolded protein response.更多还原