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。
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 Lipo- fectamine 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 trans- fected 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.
出处
《江西农业大学学报》
CAS
CSCD
北大核心
2012年第1期165-169,共5页
Acta Agriculturae Universitatis Jiangxiensis
基金
江西省科技支撑计划重点项目(2009BNA09400)
江西省自然科学基金项目(2010GZY0059)
江西农业大学校基金项目(2950)
