基于分子信标对hOGG1活性的定量分析  

Quantitative Analysis of hOGG1 Activity Based on Molecular Beacon

作　　者：刘斌[1] 羊小海[1] 王柯敏[1] 谭蔚泓[1] 

机构地区：[1]湖南大学化学生物传感与计量学国家重点实验室生物学院化学化工学院生物纳米与分子工程湖南省重点实验室,长沙410082

出　　处：《高等学校化学学报》2012年第3期486-491,共6页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:90606003);国家"八六三"计划项目(批准号:2007AA022007);湖南省杰出青年科学基金项目(批准号:08JJ1002)资助

摘　　要：设计了一种含有8-oxoG碱基的新型分子信标,结合酶促反应发展了一种非同位素标记的人8-oxoG-鸟嘌呤糖苷酶1(hOGG1)的活性分析新方法,检出限可达0.0125 U/mL.此外,该方法还可用于快速考察金属离子对酶促反应的影响和肿瘤细胞中hOGG1活性水平的定量检测.实验结果表明,该方法简单、灵敏,有望用于肿瘤样品中hOGG1活性的高通量分析和hOGG1抑制剂的筛选.Human 8-oxo-guanine DNA glycosylase 1（hOGG1）,which mainly catalyzes the excision of 8-oxoG and cleaves the AP site from DNA to sustain the genome integrity,plays an important role in many biological events including tumor development.However,available methods of hOGG1 assay based on gel electrophoresis are complicate,laborious and discontinuous.Here,we report a new method to analyze hOGG1 activity using molecular beacon containing 8-oxoG coupled with enzyme-linkage reactions,the detection limitation of this method is 0.0125 U/mL.Furthermore,the method was applied for quantification of hOGG1 activity in tumor cells and investigating the effect of metal ions on the reaction.The results demonstrate that the easy,simple and non-radioactive method has the potential to screen suitable inhibitors for hOGG1 and can be applied for high-throughput analysis of hOGG1 activity in tumor samples.

关 键 词：分子信标 人8-oxoG-鸟嘌呤糖苷酶1 实时监测 

分 类 号：O657[理学—分析化学]