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基于G-四链体的荧光共振能量转移(FRET)探针,其具有凝血酶结合适体(TBA)序列,设计用于高效荧光检测钾离子。

G quadruplex-based FRET probes with the thrombin-binding aptamer (TBA) sequence designed for the efficient fluorometric detection of the potassium ion.

作者信息

Nagatoishi Satoru, Nojima Takahiko, Galezowska Elzbieta, Juskowiak Bernard, Takenaka Shigeori

机构信息

Department of Applied Chemistry, Kyushu University, Motooka, 819-0395, Japan.

出版信息

Chembiochem. 2006 Nov;7(11):1730-7. doi: 10.1002/cbic.200600179.

Abstract

The dual-labeled oligonucleotide derivative, FAT-0, carrying 6- carboxyfluorescein (FAM) and 6-carboxytetramethylrhodamine (TAMRA) labels at the 5' and 3' termini of the thrombin-binding aptamer (TBA) sequence 5'-GGT TGG TGT GGT TGG-3', and its derivatives, FAT-n (n=3, 5, and 7) with a spacer at the 5'-end of a TBA sequence of T(m)A (m=2, 4, and 6) have been designed and synthesized. These fluorescent probes were developed for monitoring K(+) concentrations in living organisms. Circular dichroism, UV-visible absorption, and fluorescence studies revealed that all FAT-n probes could form intramolecular tetraplex structures after binding K(+). Fluorescence resonance energy transfer and quenching results are discussed taking into account dye-dye contact interactions. The relationship between the fluorescence behavior of the probes and the spacer length in FAT-n was studied in detail and is discussed.

摘要

双标记寡核苷酸衍生物FAT-0,在凝血酶结合适体(TBA)序列5'-GGT TGG TGT GGT TGG-3'的5'和3'末端带有6-羧基荧光素(FAM)和6-羧基四甲基罗丹明(TAMRA)标记,及其衍生物FAT-n(n = 3、5和7),在T(m)A(m = 2、4和6)的TBA序列的5'端带有间隔基,已被设计并合成。这些荧光探针被开发用于监测活生物体中的钾离子浓度。圆二色性、紫外可见吸收和荧光研究表明,所有FAT-n探针在结合钾离子后都能形成分子内四链体结构。考虑到染料-染料接触相互作用,讨论了荧光共振能量转移和猝灭结果。详细研究并讨论了FAT-n中探针的荧光行为与间隔基长度之间的关系。

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