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目的:筛选乙型肝炎病毒(HBV)前-前-S抗原基因启动子的结合蛋白. 方法:应用噬菌体表面展示技术,以HBV前-前-S抗原基因启动子的聚合酶链反应产物作为固相筛选分子,对噬菌体人肝细胞cDNA文库进行4轮“黏附-洗脱-扩增”的筛选过程,经噬斑的PCR扩增后,构建克隆载体,并对所筛选克隆进行DNA测序和生物信息学分析. 结果:噬菌体经富集后,从随机筛选的43个克隆中得到20 个与HBV前-前-S抗原基因启动子特异结合的阳性克隆, 包括人类SMG-1的磷脂酰肌醇3相关激酶激酶、28 S核糖体、单倍型As2A线粒体、组氨酰-tRNA合成酶、脂肪醛脱氢酶、桥粒相关蛋白、MAX相互作用蛋白1等17个已知功能基因及3个未知功能基因. 结论:用噬菌体人肝细胞cDNA文库筛选得到HBV前-前- S抗原基因启动子的结合蛋白基因,为进一步研究HBV发病机制创造了新的途径.
OBJECTIVE: To screen for the binding protein of HBV pre-pro-S antigen promoter.Methods: Polymerase chain reaction (PCR) products of HBV pre-pro-antigens gene promoter Phase screening of the phage human hepatocyte cDNA library for 4 rounds of “adhesion - elution - amplification” of the screening process, plaque PCR amplification, the construction of a cloning vector, and screening of cloned DNA sequencing and biological Informatics analysis.Results: After phage enrichment, 20 positive clones that specifically bind to HBV Pre-S antigen promoter were obtained from 43 randomly screened clones, including phosphatidylinositol of human SMG-1 17 known genes such as 3-related kinase kinase, 28 S ribosome, haploid As2A mitochondria, histidyl-tRNA synthetase, fatty aldehyde dehydrogenase, desmosome-related protein, MAX interacting protein 1 and 3 Unknown functional gene.Conclusion: The binding protein gene of HBV pre-pro-S antigen promoter was screened by the phage human hepatocyte cDNA library, which provided a new way to further study the pathogenesis of HBV.