【摘要】 AP 2α是重要的转录因子家族AP 2的成员之一.为了研究AP 2α在体内的转录调控方式及与其直接相互作用的蛋白因子,用AP 2α作为饵蛋白通过酵母双杂交系统筛选HeLa细胞的cDNA文库.筛选得到的一个阳性克隆中,经鉴定含有人TESTIN基因cDNA片段自5876bp起始的完整3′端序列.将此TESTIN片段引入GST基因融合系统成功的进行了表达.得到大小约45kD的GST TESTIN融合蛋白,用于多克隆抗体的制备.用Western blot检测TES TIN和AP 2α在HeLa细胞中的表达和定位.结果表明:TESTIN的表达主要在细胞质中,细胞核内也有少量表达;而AP 2α表达主要集中于细胞核内.它们的相互作用可能与AP 2α在细胞内分布及其调控基因转录的方式有关.更多还原

【Abstract】 AP2α is a member of an important family transcription factor AP2.In order to determine the machanism of in vivo transcriptional regulation by AP2α,a yeast twohybrid system was used to identify the interacting partners of AP2α.By using AP2α as a bait, a positive clone was obtained and sequenced.It was a cDNA fragment from 5?876?bp to 3’end of human TESTIN gene.This TESTIN fragment was inserted in frame into GST gene fusion system.A 45?kD GSTTESTIN fusion protein was produced and used for immumolige rabbit.With the polyclonal antiserum against TESTIN and monoclonal antibody against AP2α,the expression of TESTIN and AP2α in HeLa cell was determined.The result indicates that the major expression of TESTIN is in cytoplasmic extract with a little expression in nuclear extract.Whereas the expression of AP2α is concentrated in nuclear extract.Their interaction may suggest a new mechanisim to localize AP2α in the cells and to regulate the transcriptional activity of AP2α.更多还原