基于聚合酶反应和发夹型核酸适体的蛋白质荧光检测新方法  

Novel Fluorescent Method of Protein Detection Using Hairpin Nucleic AcidAptamer Based on Polymerase Reaction

作　　者：郭秋平[1] 羊小海[1] 王柯敏[1] 孟祥贤[1] 李军[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,生物医学工程中心,生命科学与技术研究院,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》2008年第1期37-40,共4页Chemical Journal of Chinese Universities

基　　金：国家重点基础研究发展规划项目(批准号:2002CB513110);国家科技攻关计划项目(批准号:2005EP090026);科技部国际合作重点项目(批准号:2003DF000039);国家自然科学基金(批准号:90606003,20475015);湖南省科技计划重点项目(批准号:0399Y1006)资助

摘　　要：设计合成了一种发夹型核酸适体（Aptamer），结合聚合酶反应建立了蛋白质荧光分析新方法．该核酸适体同时作为蛋白质配体和聚合反应模板，与靶蛋白特异结合后，其构象发生了变化，启动聚合反应，从而在未直接标记核酸适体的情况下，通过监测聚合反应进程来检测蛋白质的浓度．采用该方法检测凝血酶的线性范围为0．5～8nmoL／L，检测下限为0．5nmoL／L，为蛋白质检测提供了一种简便快速的非直接标记的荧光分析方法，有望在蛋白质组学的研究中得到广泛的应用．Aptamers are a new class of oligonucleotides generated from in vitro selection with a high affinity and specificity to targets. In this paper, a novel fluorescent method of protein detection was developed via hairpin aptamer based on polymerase reaction. The hairpin aptamer was designed as protein ligand and template of the polymerase reaction. When the aptamer was bound to the target protein, it would change to a linear strand and induce the polymerase reaction. Then protein detection was carried out by monitoring the polymerase reaction without directly labeling with the aptamer, with a linear range of 0. 5-8 nmol/Land detection limit of 0. 5 nmol/L. This proposed method has a potential advantage to design other protein probe with linear aptamer with a complex structure and can be used as a simple and general tool for protein detection.

关 键 词：核酸适体 蛋白质检测 聚合酶反应 凝血酶 

分 类 号：O657[理学—分析化学] O629.73[理学—化学]