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目的:为了探讨丙型肝炎病毒(HCV)核心(core)蛋白在肝脂肪变性中起作用的分子机制,研究了核心蛋白是否与脂肪代谢相关蛋白存在相互作用.方法:应用酵母双杂交系统3,构建HCV核心蛋白诱饵质粒,转化酵母AH109后与含文库质粒的酵母Y187进行配合,在营养缺陷培养基上进行双杂交筛选.筛选出30个克隆,对能在涂有x-α-gal的四缺营养缺陷培养基(SD/-Trp-Leu-His-Ade)上生长并变蓝的菌落,提取酵母克隆的质粒转化大肠杆菌后测序,进行生物信息学分析.结果:发现了一个与载脂蛋白A1(apoA1)同源序列,同源性99%.结论:载脂蛋白A1在酵母双杂交中能与HCV核心蛋白相互作用,推测此蛋白参与了脂质代谢紊乱,部分解释了HCV感染后普遍引起肝脂肪变性的发病机制.
Objective: To investigate the molecular mechanism of the core protein of hepatitis C virus (HCV) in hepatic steatosis and to study whether the core protein interacts with lipoprotein related proteins.Methods: By using yeast two-hybrid system, The bait plasmids of HCV core protein were constructed and transformed into yeast AH109.They were co-cultured with library plasmid Y187 and two-hybrid screening on auxotrophic medium.Among 30 clones were screened, The growth and bluish colonies of SD / -Trp-Leu-His-Ade were extracted and transformed into Escherichia coli with the yeast clones and sequenced for bioinformatics analysis.Results: The homology of protein A1 (apoA1) was 99% .Conclusion: Apolipoprotein A1 interacts with HCV core protein in yeast two-hybrid system and it is speculated that this protein is involved in lipid metabolism disorder and partially explains the effect of HCV infection The general pathogenesis of hepatic steatosis.