CytoTrap酵母双杂交筛选与乙型肝炎病毒HBx蛋白相互作用的肝细胞蛋白  

Screening of hepatocyte proteins interacting with hepatitis B virus X protein using CytoTrap yeast two-hybrid technique

作　　者：焦佰海[1] 文艳[1] 刘晓佳[1] 冯悦[1] 张阿梅[1] 刘丽[1] 夏雪山[1] 

机构地区：[1]昆明理工大学生命科学与技术学院,云南昆明650500

出　　处：《南方医科大学学报》2013年第4期486-490,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(30960346;81260248);国家科技支撑计划项目(2009BAI83B02-21;2011BAI15B01-21;2012BAI39B01)~~

摘　　要：目的筛选与乙型肝炎病毒HBx蛋白发生相互作用的肝细胞蛋白。方法构建诱饵重组载体pSos-HBx,经测序正确后将其转化酵母菌cdc25Hα中,检测其在酵母中的表达及有无自激活作用,并利用CytoTrap酵母双杂交的方法筛选与诱饵蛋白HBx相互作用的肝细胞蛋白。结果获得重组诱饵质粒pSos-HBx,验证可以在酵母中表达诱饵蛋白Sos-HBx,且诱饵质粒在此系统中没有自激活作用。利用此系统筛选出6个能与HBx相互作用的肝细胞蛋白,分别为纤连蛋白1、翻译调控肿瘤蛋白1、核受体结合蛋白IQ-WD1、软泡抑素、血清类粘蛋白1、二硫化物异构酶家族A3。结论成功克隆出乙型肝炎病毒HBx蛋白的结合蛋白,为进一步研究HBx蛋白在肝炎或肝癌过程中发挥的作用提供了新线索。Objective To screen the hepatocyte proteins that interact with hepatitis B virus X protein（HBx）.Methods The recombinant plasmid pSos-HBx was constructed by inserting Sos-HBx fragment into the bait vector,and after sequence verification the plasmid was transformed into competent yeast cells.The expression and self-activation of Sos-HBx protein was detected in the yeast cells.The hepatocyte proteins interacting with the bait protein was screened with CytoTrap yeast two-hybrid technique.Results The reconstructed plasmid harboring HBx gene expressed Sos-HBx protein in the yeast cells without self-activation of the protein.CytoTrap yeast two-hybrid system identified 6 hepatocyte proteins that interacted with HBx,including fibronectin 1,translationally controlled tumor protein,IQ motif and WD repeats 1,follistatin,orosomucoid 1, and disulfide isomerase family A member 3.Conclusion Six HBx-binding hepatocyte proteins have been identified using the CytoTrap yeast two-hybrid system,which provides clues for further investigation of the role of HBx protein in hepatitis and liver cancer.

关 键 词：HBX蛋白 CytoTrap酵母双杂交 蛋白质相互作用 

分 类 号：R373[医药卫生—病原生物学]