催化发夹组装用于碱基切除修复酶活性检测研究  

An Enzyme-Free Signal Amplification Platform Based on Catalytic Hairpin Self-Assembly for Detection of Endonuclease IV Activity

作　　者：王涛 汤建新 李青 WANG Tao;TANG Jianxin;LI Qing(College of Life Sciences and Chemistry,Hunan University of Technology,Zhuzhou Hunan 412007,China)

机构地区：[1]湖南工业大学生命科学与化学学院

出　　处：《包装学报》2019年第3期24-29,共6页Packaging Journal

基　　金：国家自然科学基金资助项目(21705042);中国包装联合会“绿色包装与安全”专项研究基金资助项目(2017ZBLY14)

摘　　要：碱基切除修复酶在DNA损伤修复过程中具有重要作用,且其检测与癌症等疾病的诊断相关。传统的碱基切除修复酶检测方法操作复杂、灵敏度低,且仅对酶的浓度进行定量分析而非酶的活性。为此,建立了基于催化发夹组装介导信号放大用于核酸内切酶IV(Endo IV)活性的检测方法。该方法利用Endo IV的活性作用于底物探针,将引发序列释放而引发催化发夹自组装信号放大,以实现Endo IV的活性检测分析。通过荧光检测实验可知,该方法检测下限为3.7×10-7 U/mL,可选择性地对Endo IV的活性进行检测,是一种设计简单、操作简便、灵敏度高的碱基切除修复酶活性检测方法。Base excision repair enzymes played an important role in DNA damage repair,and their detection was related to cancer and disease diagnosis.The traditional detection of base excision repair enzymes was complex and lowsensitive,which only quantitatively analyzed the concentration of enzymes,rather than enzyme activity detection.A signal amplification platform based on catalytic hairpin self-assembly was established to detect endonuclease IV activity in base excision repair enzymes.This method was based on the activation of endonuclease IV acting on the substrate probe and releasing the initiation sequence,which caused the amplification of self-assembly signal of catalytic hairpin to realize the detection and analysis of endonuclease IV activity.According to the fluorescence detection experiment,the detection lower limit of this method was 3.7×10-7 U/mL,and Endo IV activity could be selectively detected.Therefore,this method was simple in design and operation and high in sensitivity.

关 键 词：催化发夹组装 碱基切除修复酶 酶活性检测 无酶信号放大 核酸内切酶IV 

分 类 号：F274[经济管理—企业管理] F224[经济管理—国民经济]