基于富G序列的无标记荧光传感器检测单链核酸

任林娇 薛梦晓 陈青华 魏铭航 张培 秦自瑞 闫艳霞 姜利英

引用本文: 任林娇, 薛梦晓, 陈青华, 魏铭航, 张培, 秦自瑞, 闫艳霞, 姜利英. 基于富G序列的无标记荧光传感器检测单链核酸[J]. 分析化学, 2023, 51(6): 962-971. doi: 10.19756/j.issn.0253-3820.221546 shu
Citation:  REN Lin-Jiao,  XUE Meng-Xiao,  CHEN Qing-Hua,  WEI Ming-Hang,  ZHANG Pei,  QIN Zi-Rui,  YAN Yan-Xia,  JIANG Li-Ying. Label-free Fluorescent Sensor for Detection of Single-stranded Nucleic Acids Based on G-rich Sequence[J]. Chinese Journal of Analytical Chemistry, 2023, 51(6): 962-971. doi: 10.19756/j.issn.0253-3820.221546 shu

基于富G序列的无标记荧光传感器检测单链核酸

    通讯作者: 陈青华,E-mail:2006007@zzuli.edu.cn; 姜利英,E-mail:jiangliying@zzuli.edu.cn
  • 基金项目:

    国家自然科学基金项目(Nos.62073299,12004348,61801436)、中原科技创新领军人才项目(No.224200510026)和河南省科技攻关项目(Nos.212102210196,212102210002,212102210278,212102210008)资助。

摘要: 具有特定构象的富G序列(如G-四链体和G-三链体)与荧光染料相互作用可有效增强其荧光信号强度,被广泛应用于构建无标记荧光生物传感。本研究以硫磺素T(Thioflavin T,ThT)为荧光染料,构建了两种基于富G序列的无标记荧光传感器,用于检测阿尔兹海默病标志物β-淀粉样蛋白(β-Amyloid protein,Aβ)的基因序列。实验结果表明,分子发夹茎长为4个碱基时,富G序列以G-三链体的结构存在,以此构建的G-三链体传感器的输出信号随Aβ基因浓度增加而降低,线性检测范围为1~100 nmol/L,检出限为0.3 nmol/L(S/N=3)。分子发夹茎长为8个碱基且5'端添加碱基AATT时,其与Aβ基因结合后,富G序列多以G-四链体结构存在,以此构建的G-四链体传感器的输出信号随Aβ基因浓度增加而增强,线性检测范围为0.1~100 nmol/L,检出限为0.04 nmol/L (S/N=3)。两种传感器制备过程相似但检测原理不同,为富G序列的深入研究与应用提供了参考,同时为单链核酸的无标记荧光检测提供了新的思路。

English


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  • 收稿日期:  2022-11-03
  • 修回日期:  2023-04-03
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