【摘要】 基于双链DNA结合染料能特异嵌入双链DNA发出荧光的原理,发展了一种实时检测DNA聚合酶活性的简便方法.在检测过程中,聚合酶的聚合反应进程被实时转换为荧光信号,通过监测荧光强度的变化实时检测聚合酶的活性及药物对聚合酶活性的影响.该方法不需要对DNA进行放射性同位素标记和荧光标记,也不需要聚丙烯酰胺凝胶电泳和聚合酶链式反应,是一种简便、快速的聚合酶活性实时检测新方法,为研究抗肿瘤药物对聚合酶活性的影响提供了一种简捷方法,也将为相关疾病诊治和药物筛选提供一种新的思路.更多还原

【Abstract】 Traditionally,polymerase is assayed by denaturing gel electrophoresis and autoradiography,which are complex and discontinuous. Based on the fact that the dye SYBR Green I can intercalate double-stranded DNA to emit fluorescence,we report a new fluorescence-based DNA polymerase assay that can translate the polymerization process into a fluorescent signal in real-time. Comparatively,the method is simple,convenient,less time consuming and inexpensive without the need of gel electrophoresis,radial labeling,fluorescent labeling and polymerase chain reaction (PCR). Based on this assay,the influence of anti-tumour drugs on the activity of DNA polymerase is investigated. This method will provide a potential in diagnosis and therapy of disease and selecting drugs.更多还原