Strand-invading linear probe combined with unmodified PNA

IF 3.5 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Bioorganic & Medicinal Chemistry Pub Date : 2016-09-15 Epub Date: 2016-06-29 DOI:10.1016/j.bmc.2016.06.055

Hiroyuki Asanuma, Rie Niwa, Mariko Akahane, Keiji Murayama, Hiromu Kashida, Yukiko Kamiya

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引用次数: 6

Abstract

Efficient strand invasion by a linear probe to fluorescently label double-stranded DNA has been implemented by employing a probe and unmodified PNA. As a fluorophore, we utilized ethynylperylene. Multiple ethynylperylene residues were incorporated into the DNA probe via a d-threoninol scaffold. The ethynylperylene did not significantly disrupt hybridization with complementary DNA. The linear probe self-quenched in the absence of target DNA and did not hybridize with PNA. A gel-shift assay revealed that linear probe and PNA combination invaded the central region of double-stranded DNA upon heat-shock treatment to form a double duplex. To further suppress the background emission and increase the stability of the probe/DNA duplex, a probe containing anthraquinones as well as ethynylperylene was synthesized. This probe and PNA invader pair detected an internal sequence in a double-stranded DNA with high sensitivity when heat shock treatment was used. The probe and PNA pair was able to invade at the terminus of a long double-stranded DNA at 40 °C at 100 mM NaCl concentration.

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链入侵线性探针结合未修饰的PNA

利用探针和未修饰的PNA实现了线性探针对双链DNA荧光标记的有效入侵。作为荧光团，我们使用乙炔-苝。多个乙基苝残基通过d-苏氨酸醇支架并入DNA探针中。乙炔对与互补DNA的杂交没有明显的破坏作用。在没有目标DNA的情况下，线性探针自猝灭，不能与PNA杂交。凝胶移位实验显示，经热休克处理后，线性探针和PNA组合侵入双链DNA的中心区域，形成双双工。为了进一步抑制背景辐射，提高探针/DNA双链的稳定性，合成了一种含有蒽醌和乙基苝的探针。该探针和PNA入侵对在热休克处理时以高灵敏度检测双链DNA的内部序列。探针和PNA对能够在40°C和100 mM NaCl浓度下侵入长双链DNA的末端。

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来源期刊

Bioorganic & Medicinal Chemistry 医学-生化与分子生物学

CiteScore

6.80

自引率

2.90%

发文量

413

审稿时长

17 days

期刊介绍： Bioorganic & Medicinal Chemistry provides an international forum for the publication of full original research papers and critical reviews on molecular interactions in key biological targets such as receptors, channels, enzymes, nucleotides, lipids and saccharides. The aim of the journal is to promote a better understanding at the molecular level of life processes, and living organisms, as well as the interaction of these with chemical agents. A special feature will be that colour illustrations will be reproduced at no charge to the author, provided that the Editor agrees that colour is essential to the information content of the illustration in question.