通过以转录为媒介的扩增和杂交保护实验来测量端粒酶活性的新方法  

New Method to Measure Telomerase Activity by Transcription-Mediated Amplification and Hybridization Protection Assay

作　　者：聂卉[1] 霍丽云[2] 

机构地区：[1]山东大学,山东济南250100 [2]山东省教育学院生化系,山东济南250013

出　　处：《山东教育学院学报》2002年第2期84-86,共3页Journal of Shandong Education Institute

摘　　要：利用TMA HPA技术 ,可以从 1或 2 5个细胞当量中探测出端粒酶的活性。其探测范围大于传统的TRAP技术。并且 ,当样品抽提物被稀释到≤ 1μg蛋白时 ,未用酚—氯仿处理过的TMA HPA给出的信号与用其处理过而给出的信号是相同的。说明在低浓度蛋白的临床样品中 ,该技术几乎不受TRAP抑制因子的作用。当含有 1μg或 0 1μg蛋白时 ,在 33个从肝脏中提取出的组织样品中 ,有 2 7个给出阳性信号 ,阳性比率为 81 8%,说明TMA HPA的灵敏度较高。TMA/HPA could detect the telomerase activity of 1 or 2.5 cell equivalents extracted from cell lines.The detection limit of TMA/HPA was equivalent or better to that of conventional TRAP.Morever,when extracts were diluted and of protein was used,TMA/HPA signals without phenol-chloroform treatment were equivalent to those with phenol-chloroform treatment.This demonstrated that TMA/HPA minimizes the influence of inhibitors in diluted clinical samples with low protein concentration.Tissue samples from liver were used to verify the detection limit of TMA/HPA. Results confirmed the sensitivity of TMA/HPA.The positive rats was 81.8%(27 of 33)when either 1 or 0.1 of protein was used.

关 键 词：转录 扩增 杂交保护实验 端粒酶 TRAP技术 TMA/HPA技术 染色体复制 

分 类 号：Q55[生物学—生物化学]