酵母双杂合体系研究丙型肝炎病毒进入肝细胞的受体

Study on the receptor of HCV entry into the hepatocyte by yeast two hybrid system

目的 :用酵母双杂合体系筛选肝细胞cDNA文库 ,寻找与丙型肝炎 (丙肝 )病毒 (hepatitisCvirus ,HCV)胞膜蛋白 2 (E2 )相作用蛋白的基因 ,探讨HCV与肝细胞的相互作用 ,寻找HCV进入细胞的受体。方法 :以HCVE2蛋白胞外区片段 ,构建酵母双杂合体系中“饵”载体 ,从成人肝细胞cDNA文库筛选与HCVE2相作用蛋白的基因。结果 :筛出的阳性克隆基因序列分析后 ,经GenBank查询 ,与人的apoCⅢ同源 ,同源性为 98%。 结论 :HCVE2蛋白能与人的apoCⅢ在酵母双杂合体系中结合 。

Objective: To select the gene from human liver cDNA library which can transform the protein that can interact with HCV E2 protein by yeast two hybrid system and look for the receptor by which HCV enters the host cell. Methods: Hepatitis C virus E2 gene without transmembrane domain was used to construct the “bait plasmid” to look for the gene from the cDNA library, which expressed the protein that could interact with the HCV E2 protein. Results: After sequencing the positive clone was provided to the GenBank. It showed that the cDNA was homologous to the human apolipoprotein CⅢ (identities 98%). Conclusion: HCV E2 protein can interact with human apoC Ⅲ in the yeast two hybrid system. But the relationship of the HCV vLDL and the HCV entering the host is still being studied.