高等学校化学学报 ›› 2012, Vol. 33 ›› Issue (07): 1407-1412.doi: 10.3969/j.issn.0251-0790.2012.07.008

• 分析化学 • 上一篇    下一篇

一种高效灵敏的DNA荧光探针的合成及应用

吴彤, 彭孝军, 胡明明, 刘飞, 樊江莉   

  1. 大连理工大学精细化工国家重点实验室, 大连 116024
  • 收稿日期:2011-11-21 出版日期:2012-07-10 发布日期:2012-07-10
  • 通讯作者: 樊江莉, 女, 博士, 副教授, 主要从事荧光染料和荧光探针研究. E-mail: fanjl@dlut.edu.cn E-mail:fanjl@dlut.edu.cn
  • 基金资助:

    国家自然科学基金(批准号: 21136002, 21076032, 20923006)资助.

Synthesis and Application of an Efficient and Sensitive DNA Fluorescent Probe

WU Tong, PENG Xiao-Jun, HU Ming-Ming, LIU Fei, FAN Jiang-Li   

  1. State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024, China
  • Received:2011-11-21 Online:2012-07-10 Published:2012-07-10

摘要: 设计合成了一种新型噻唑橙二聚体荧光染料Bi-TO3, 采用荧光发射光谱、 圆二色光谱及活细胞荧光成像等方法研究了其与DNA的相互作用. 在10 mmol/L Tris-HCl缓冲液(pH=7.4)中, Bi-TO3的固有荧光极弱, 量子产率小于0.001%; 与小牛胸腺DNA结合后, 其荧光可显著增强约950倍, 但对RNA和蛋白等生物大分子及黏度等环境因素则无明显响应. 紫外吸收光谱及圆二色光谱滴定实验表明, Bi-TO3以小沟结合模式与DNA作用, 且对AT序列有选择性. 实验结果表明, 在缓冲溶液中Bi-TO3的荧光增强信号与低浓度范围的poly(dA-dT)2仍呈良好的线性关系, 检出限为13.3 ng/mL, 灵敏较度高; 且Bi-TO3可在较低浓度范围(6~12 μmol/L)内应用于活细胞荧光成像.

关键词: 噻唑橙二聚体, 荧光探针, DNA定量检测, 活细胞荧光成像

Abstract: A novel thiazole orange dimmer derivative Bi-TO3 was developed. The interaction of Bi-TO3 with DNA was investigated through the absorption, emission, circular dichroism spectroscopy and live-cell imaging experiments. As a specific probe for DNA, Bi-TO3 itself was almost non-fluorescent(quantum yield<0.001%) in buffer. Upon binding to DNA, the fluorescence of Bi-TO3 increased approximately 950-fold, even without interferences of responding to RNA, BSA, viscosity, etc. Absorption and circular dichroism spectra showed that Bi-TO3 associated with DNA in a minor groove binding mode. The emission detection limit was 13.3 ng/mL, which manifested that Bi-TO3 could detect trace DNA in solution. Moreover, Bi-TO3 could be applied for live-cell fluorescence imaging with relatively low concentrations of 6-12 μmol/L.

Key words: Thiazole orange dimmer, Fluorescent probe, DNA quantitative detection, Live-cell fluorescence imaging

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