Objective To screen and identify proteins that intact with Hepatitis B virus(HBV) X protein by means of T7-phage display system. Methods gene was cloned and expressed by prokaryotic expression,the purified protein was used as a selected protein to biopan the T7 select human liver cDNA library, the selected positive clones were identified using DNA sequencing and searched for homologous. Results X protein was expressed and purified,after 4 rounds of biopanning,52 positive clones were selected. The inserted fragments were amplified by PCR using T7 primer. The products were purified for sequencing,7 proteins were identified to interact with X protein by homologous analysis. Conclusion T7 select phage display system is a convenient and rapid method for screening binding proteins. The result will be useful for further studying the pathogenic mechanism of HBV.%目的 利用T7噬菌体展示技术筛选与乙型肝炎病毒X蛋白相互作用的蛋白.方法 构建X蛋白的原核表达载体,表达并纯化X蛋白,以X蛋白作为靶蛋白,应用T7噬菌体展示技术对人肝细胞cDNA文库进行筛选,对筛选到的阳性克隆进行DNA序列分析及同源性研究.结果 表达并纯化了X蛋白,经过4轮筛选后,选择52个阳性克隆,采用T7特异性引物扩增插入片段,回收PCR产物进行测序.经过同源性分析,确定了7个与乙型肝炎X蛋白相互作用的蛋白.结论 T7噬菌体展示筛选是1种快速、简单的筛选蛋白质相互作用的工具,筛选的与X蛋白相互作用蛋白为进一步研究乙型肝炎病毒的致病机制奠定了基础.
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