用酵母单杂交系统筛选人IL-2Rα基因NIRS元件结合蛋白的cDNA  

Screening of cDNA encoding for human interleukin-2 receptor alpha gene NIRS element binding proteins by using yeast one-hybrid system

作　　者：盛德乔[1] 陆瑜[1] 李宏帆[1] 吴宁华[1] 沈珝琲[1] 

机构地区：[1]中国医学科学院中国协和医科大学基础医学研究所医学分子生物学国家重点实验室,北京100005

出　　处：《基础医学与临床》2005年第3期214-218,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(30270312)

摘　　要：目的筛选与白细胞介素2受体(IL2R)α基因NIRS元件相互作用的结合蛋白。方法采用酵母单杂交体系从HTLV1转化的人外周血淋巴细胞MATCHMAKERcDNA文库中筛选与NIRS相互作用的蛋白。结果经过筛选并排除假阳性后有9个阳性克隆仍保持His+表型和βgal活性;测序后分析发现它们分别属于4个不同蛋白的cDNA克隆。其中一个是已知的反式作用因子Ku抗原,另一个是RNA聚合酶Ⅰ的转录终止因子。它们的C末端分别含有SAP和SANTDNA结合结构域。结论在Jurkat细胞中存在与NIRS元件相互作用的结合蛋白,相关的研究正在进行中。Objective To screen the binding proteins of Human Interleukin-2 receptor α Chain NIRS element. Methods Yeast one-hybrid system was used to screen NIRS binding proteins in HTLV-1 transformed Jurkat cell cDNA fusion library. Results There are nine clones have the His + epitope and β-gal activity after the secondary screening. The sequencing results indicate that they are belonging to 4 different proteins. Among them, a known trans-acting factor Ku antigen and transcription termination factor of RNA polymerase I (TTF1) contain SAP domain and SANT domain, respectively. They are DNA binding domains. Conclusion Four candidate clones encoding for NIRS binding proteins were obtained from Jurkat cells . The regulation mechanism of these candidate proteins in IL-2R α gene is undergoing.

关 键 词：NIRS 结合蛋白 人IL-2 筛选 Ra基因 杂交系统 酵母 白细胞介素2受体 人外周血淋巴细胞 DNA结合结构域 Jurkat细胞 相互作用 HTLV-1 cDNA文库 cDNA克隆 反式作用因子 RNA聚合酶 阳性克隆 Ku抗原 杂交体 gal 假阳性 SAP C末端 

分 类 号：Q781[生物学—分子生物学]