中文摘要：

目的：应用酵母双杂交系统从人正常胃黏膜细胞的cDNA文库中筛选E2F1转录因子的相互作用蛋白.方法：以pGBKT7-E2F1为诱饵质粒筛选人正常胃黏膜细胞的cDNA文库,得到阳性克隆,反复验证,并对验证后的阳性克隆进行测序及同源性分析.结果：从人正常胃黏膜细胞的cDNA文库中筛选得到20个阳性克隆,经测序和同源性分析,筛除假阳性克隆,最终得到18个不同的候选基因序列.其中,17个为可知基因序列,它们分别是：组织蛋白酶B、SIVA1、干扰素调节因子7、鸟苷酸激酶1、ATP酶抑制因子1、核糖体蛋白SA、纤溶酶原激活物抑制剂1 mRNA结合蛋白、精氨琥珀酸合酶1、卵泡抑素类似物3、金属硫蛋白2A、内质网钙结合蛋白1、WNT1可诱导信号通路蛋白2、CDC42 效应蛋白1、可溶性半乳糖凝集素结合蛋白1、中胚层发育候选2、外切酶体成分7和含EGF腓骨蛋白样胞外基质蛋白 1,尚有一未知基因片段.结论：应用酵母双杂交系统成功筛选出18种E2F1相互作用蛋白,为进一步探讨E2F1影响胃癌细胞生物学行为的分子机制奠定了基础.

英文摘要：

AIM：To screen the proteins which interact with E2F1 transcription factor by yeast two-hybrid system in human gastric mucous cells. METHODS：The cDNA library of human gastric mucous cells was screened with pGBKT7-E2F1 as a bait plasmid by yeast two-hybrid system. The His＋/Leu＋/Trp＋/Ade＋/LacZ＋ yeast clones were obtained. Moreover, the inserted fragments of identified positive clones were sequenced and analyzed by bioinformatic methods. RESULTS：Twenty His＋/Leu＋/Trp＋/Ade＋/LacZ＋ yeast clones were obtained through screening the cDNA library of human gastric mucous cells with pGBKT7-E2F1 as a bait plasmid. Eighteen sequences of different candidate genes were confirmed by BLAST analysis in NCBI including cathepsin B, SIVA1, metallothionein 2A, ribosomal protein SA, guanylate kinase 1, ATPase inhibitory factor 1, serpine 1 mRNA binding protein, argininosuccinate synthase 1, mesoderm development candidate 2, WNT1 inducible signaling pathway protein 2, reticulocalbin 1, CDC42 effector protein 1, exosome component 7, EGF-containing fibulin-like extracellular matrix protein 1, follistatin-like 3, interferon regulatory factor 7, lectin galactoside-binding soluble 1 and another unknown gene sequence. CONCLUSION：Eighteen proteins interacting with E2F1 have been successfully screened from the cDNA library of human gastric mucous cells by yeast two-hybrid system, which will be helpful for further investigating the role of E2F1 in the biological behaviors of gastric carcinoma cells.