基于T-Hg2+-T及G四聚体自身熄灭能力的“Turnon”型单标记DNA荧光探针用于碘离子的检测

利用G四聚体可以熄灭荧光的特性以及T-Hg2+-T的特殊结构, 发展了一种简便的"Turn on"型碘离子检测新方法. 设计了一条5'端标有荧光基团的富T序列, 3'端采用能形成G四聚体的富G序列代替传统的熄灭基团. 加入汞离子后, 富T序列形成T-Hg2+-T机构发生折叠, G四聚体靠近荧光基团, 发生光诱导电子转移, 使荧光被熄灭. 若加入碘离子, 碘离子会与汞离子形成较稳定的配合物, 汞离子从DNA上被竞争下来, 探针的荧光得以恢复, 且荧光强度与50～500 nmol/L的碘离子呈良好线性关系, 检出限为30 nmol/L. 本方法选择性好, 10倍于碘离子浓度的其它常见阴离子干扰较小. 检测自来水样中碘离子的回收率为92%～109%, 相对标准偏差RSD<4%(n=4).

Single-labeled“Turn on” Fluorescent Oligonucleotide Probe for Iodide Detection Based on T-Hg2+-T and Inherent Quenching Ability of G-quadruplex

A simple and "Turn on" type iodide anion assay method was developed, based on the inherent quenching ability of G-quadruplex and fidelity of the "thymine-Hg2+-thymine" binding motif. A T-rich sequence was labeled with 6-carboxyfluorescein(FAM) at its 5'-end, nearby the 3'-end was a G-rich sequence which could form G-quadruplex structure instead of traditional quenchers. Upon addition of Hg2+, T-rich sequence folded into a hairpin structure which led the G-quadruplex near to the FAM, and the FAM was quenched by the G-quadruplex owning to photoinduced electron transfer between the dye and the G-quadruplex. After the addition of iodide, the fluorescence of FAM recovered considering that iodide could bind with Hg2+. The novel method for the determination of iodide was developed in linear range of 50—500 nmol/L, with the detection limit(3σ) of 30 nmol/L. This proposed method was highly selective and other anions have no interfering effects on the determination, which was successfully applied for analysis of real samples with the recovery from 92%—109%, and the RSD<4%(n=4).