PC-1分子转录激活功能研究

The Study of The Transcriptional Activation Activity of PC-1

PC 1基因是在人前列腺癌细胞中克隆的新基因 ,表达水平随前列腺癌恶性程度增加而升高 ,其表达产物具有转录因子的一些特征 .为研究PC 1分子的转录激活功能 ,首先应用酵母双杂交系统将PC 1全长以及不同区段的cDNA克隆到表达载体pAS2 1中 ,然后分别转化酵母细胞株CG 194 5 .lacZ和His3报告基因激活的检测结果表明 ,该分子具有转录激活活性并将该活性定位于N端的 4 6个氨基酸区域 .此外 ,将PC 1分子不同区段的cDNA分别克隆至表达载体pZHO1中 ,将它们与报告基因质粒pTRE luc共转染哺乳动物细胞COS7和C4 2 ,Firefly荧光素酶相对活性的检测结果表明 ,该分子N端的 4

The gene PC-1 was identified as a novel gene that expressed higher in the much more malignant human prostate cancer cell lines and its translation product meets some points of transcription factors. In order to study the transcriptional activation activity of PC-1, firstly, the yeast two-hybrid system was used and the whole length as well as various regions of PC-1cDNA were cloned into the expression vector pAS2-1 and transfected into the yeast cell line CG-1945 respectively, the result of the reporter genes lacZ and His3 activation assay showed that PC-1 had transcriptional activation activity and this activity was mapped within its N terminal 46 amino acids. Then, the various regions of PC-1 cDNA were cloned into the expression vector pZHO1 and cotransfected with the pTRE-luc plasmid of the reporter gene into COS7 and C4-2 mammalian cells, the 46 amino acids in the N terminal of PC-I were found to have transcriptional activation activity by the firefly relative luciferase activity assay.