DNA酶的设计、选择及催化动力学研究  

Study on the Design and Selection of DNAzymes and the Kinetics of their Catalytic Reactions

作　　者：李英[1] 余晓冬[1] 徐静娟[1] 陈洪渊[1,2] 

机构地区：[1]南京大学化学化工学院生命分析化学国家重点实验室,南京210093 [2]中国科学院

出　　处：《化学通报》2013年第7期617-623,共7页Chemistry

基　　金：国家自然科学基金重大项目课题(21190044);国家自然科学基金创新研究群体项目(21121091)资助;国家重大科学仪器设备开发专项子课题(2011YQ17006711);973计划课题(2011CB911003)

摘　　要：设计了12条能与血晶素相互作用形成DNA酶的寡核苷酸链,并研究了它们催化4种不同底物[2,2-联氮基双(3-乙基苯并噻唑林-6-磺酸)二铵盐、2,7-二氯二氢荧光素二乙酯、酪胺、四甲基联苯胺]的催化活性及动力学过程。结果表明,G-四分体中碱基G形成的平面结构的层数、可以形成类发夹结构的互补DNA短链、G-四分体侧环上碱基的个数以及类发夹结构连接臂上碱基的个数等因素均影响血晶素与DNA的相互作用。除此之外,DNA酶的催化活性不仅与血晶素和DNA的结合常数有关,同时也受底物分子结构的影响。在所设计的12种DNA酶中,2,2-联氮基双(3-乙基苯并噻唑林-6-磺酸)二铵盐是理想的比色测定底物,而酪胺是理想的荧光测定底物。In this paper, twelve nucleic acids were designed as aptamers to form DNAzymes with hemin. Four compounds, ABTS, H2DCFDA, tyramine and TMB were selected as substrates to be catalyzed by these DNAzymes. The results showed that the number of G-strand layers, the hairpin-like DNA duplex, the number of the nucleic acid bases in the lateral loops and spacers of hairpin-like DNA duplex all affect the hemin-binding affinity with aptamers. Besides, the catalytic activity of DNAzymes was not only determined by hemin-binding affinity but also affected by the spatial structure of substrate. For these DNAzymes, ABTS and tyramine are a good substrate for colorimetric determination and fluorimetric determination, respectively.

关 键 词：G-四分体 血晶素 DNA酶 

分 类 号：Q55[生物学—生物化学]