一种长时稳定的铅离子响应型脱氧核酶的合成与表征  

Synthesis and Characterization of a Long-term Stable Lead-dependent DNAzyme

作　　者：陈词 蒯海岚 李林羚 胡霞 杨基峰 张松柏 CHEN Ci;KUAI Hai-an;LI Lin-ing;HU Xia;YANG Ji-feng;ZHANG Song-bai(Hunan Provincial Key Laboratory of Water Treatment Functional Materials,Hunan Province Engineering Research Center of Electroplating Wastewater Reuse Technology,College of Chemistry and Materials Engineering,Hunan University of Arts and Science,Changde 415000,China)

机构地区：[1]湖南文理学院,化学与材料工程学院,水处理功能材料湖南省重点实验室,电镀废水回用技术湖南省工程研究中心,湖南常德415000

出　　处：《分析测试学报》2024年第1期191-199,共9页Journal of Instrumental Analysis

基　　金：湖南省自然科学基金项目(2022JJ30419,2022JJ30418);湖南省教育厅科学研究项目(23A0503,19B384);常德市科技创新项目(CDKJJ20220522);湖南文理学院校级科研启动项目(21BSQD14,22BSQD21)。

摘　　要：脱氧核酶作为一类具有酶活性的功能核酸分子,以其不同于蛋白酶和核酶的结构优势而被广泛应用于生物传感等分析检测领域。但核酸结构易受生物酶降解,故而限制了脱氧核酶在该领域应用研究中的推进。化学修饰的方法可改善该问题,但仍存在修饰繁琐且价格昂贵等弊端。该文基于核酸结构改造,利用环形核酸结构的耐酶切能力,设计合成了一种环状结构的铅离子响应型脱氧核酶,并对其性能进行了表征。研究结果表明该环形脱氧核酶在外切酶样、自来水样以及75%胎牛血清(FBS)中具有比传统单链脱氧核酶更好的结构稳定性,其中在75%胎牛血清中稳定存在的时长达6h。此外,环形结构的引入也增强了脱氧核酶与底物序列的相互作用,进而影响基于环形脱氧核酶的生物传感器的信号表达,这一特性有望为脱氧核酶生物传感器的构建提供新思路。As a type of functional nucleic acid with enzymatic activity,DNAzymes are widely used in analytical fields such as biosensing due to their structural advantages that are different from proteas-es and ribozymes.However,the structure of DNAzyme is susceptible to degradation by biological en-zymes,which slows down the speed of such biosensor development.Although chemical modification can be used to resolve the problem,this strategy has disadvantages,such as being costly and hard to synthesize.Considering this,we choose a new strategy from the sights of structure transformation to resolve this problem.Then,we design and synthesize a kind of circular DNAzyme by utilizing the en-zyme-resistance properties of circular nucleic acid structure.The research results show that the circu-lar DNAzyme has better structural stability than traditional single-stranded DNAzymes in exonucle-ase,tap water,and 75%fetal bovine serum(FBS),and the duration is up to 6 h in 75%FBS.In ad-dition,introducing a circular structure will also enhance the interaction between DNAzymes and sub-strate sequences,thereby affecting the signal expression of biosensors based on circular DNAzymes.

关 键 词：环形脱氧核酶 环形核酸 脱氧核酶 铅离子检测 生物传感器 

分 类 号：O657.3[理学—分析化学] O614.433[理学—化学]