酵母单杂交筛选人食管癌细胞系SHEEC中NF-kappa B元件结合蛋白

背景与目的：从食管癌细胞cDNA文库中筛选能够特异性结合NF-kappa B核心元件的蛋白质分子，探讨NGAL基因的表达调控机制，探寻NGAL基因在食管上皮细胞癌变中过表达的原因。材料与方法：应用酵母单杂交系统，以三重NF-kappa B元件寡聚核苷酸串为诱饵，对人食管癌细胞系SHEEC cDNA酵母杂交文库进行筛选，应用DNA测序及生物信息学技术对筛选克隆进行分析。结果：共获得30个阳性克隆，包括4种编码蛋白，2个hypothetical protein，2种线粒体基因，3类非编码RNA等。结论：应用酵母单杂交手段筛选出食管癌细胞NF-kappa B核心元件结合蛋白，但只是一种初步的实验，还需进一步研究证实。

Screening NF-kappa B Element Binding Protein of Esophageal Carcinoma Cell Line SHEEC With Yeast One Hybrid System

BACKGROUND&AIM: This study was to screen an esophageal cancer cell line(SHEEC)cDNA library for the cDNA that expressed a protein that can specifically bind the NF_kappa B element,for further understanding the expression and regulation mechanism of the NGAL(neutrophil gelatinase_associated lipocalin)gene,and the course for the over_expression of the NGAL gene in the esophageal epithelial squamous carcinoma. MATERAL AND METHODS: A tandem three oligo_nucleotides of NF_kappa B was employed as a bait in a yeast one hybrid system to screen the cDNA library of the human esophageal carcinoma cell line(SHEEC).The cDNA clones obtained were sequenced and analyzed with bioinformatics tools. RESULTS: Among the thirty clones obtained,four types of clones could encode proteins,two clones were hypothetical proteins,two types of clones were mitochondrial genes,and three types of clones were non_encoding sequences. CONCLUSION: Using the yeast one hybrid system to screen the cDNA library for the NF_kappa B core element is a preliminary work that needs confirmation by further study.