摘要
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.
AIM: Previous study suggested that the core protein of hepa-titis C virus (HCV) may alter lipid metabolism. To under-stand the role of this protein in HCV pathogenesis,the au-thors investigated the interaction of HCV core protein withother cellular protein.METHODS: Using the yeast two hybrid system 3, the au-thors constructed bait plasmid with the the core proteingene of hepatitis C virus. After confirmed that hepatitis Cvirus core protein could firmly expressed in AH109 yeaststrains, yeast two hybrid assay was performed by matingAH109 with Y187 that transformed with liver cDNA libraryplasmids and then plated on quadruple dropout (QDO) me-dium and assayed for x-α -gal activity. Sequencing analysis ofthe genes of library plasmids was conducted in yeast coloniesthat could grow on QDO medium covered with x-α -gal.RESUITS: Among the 30 positive colonies, the authors founda homologous gene – apoA1, one of the protein compo-nents of high-density lipoprotein.CONCLUSION: The core protein of HCV can interact withapoA1, which may partly explain the liver steatosis occur-ring in hepatitis C.
出处
《世界华人消化杂志》
CAS
2002年第9期1018-1021,共4页
World Chinese Journal of Digestology
