Gene Detection by Light-to-electric Converting Intercalator and Its Application to DNA

光电转换嵌入剂基因检测及其在DNA中的应用

基本信息

批准号：
12305054
负责人：
TAKAGI Makoto
金额：
$ 15.33万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12305054/
关键词：
photocurrent gene detection DNA chip intercalator ruthenium complex threading intercalation

项目摘要

In this research we developed a novel detection system of DNA duplex using a light-to-electric conversion. The principle of the detection of DNA duplex formation is as follows. 1) The gold electrode modified by single stranded DNA was prepared. 2) When double stranded DNA is formed, ruthenium complex will intercalate and will be concentrated on the electrode surface. 3) Then the visible light is irradiated to the electrode, photocurrent caused by excitation of ruthenium complex will be observed.We used a polypyridyl ruthenium complex carrying DNA intercalating ligand for DNA detection. Ru(bpy)_3 was used as a control compound. Ru(bpy)_2dppz was used as a ruthenium complex carrying a DNA intercalating part. Furthermore, we synthesized NDI-Ru_2 as a threading intercalator carrying ruthenium complex. This type of compound has a smaller dissociation rate from DNA duplex, because it has threading intercalating part connected to two bulky groupsA DNA-immobilized gold electrode was dipped into the solution containing the light-to-electric converting intercalator (Ru(bpy)_3, Ru(bpy)_2dppz, NDI-Ru_2). When the visible light was irradiated to the surface of the electrode, continuous photocurrent occurred in the presence of tertiary amine as an electron donor. The amount of photocurrent increased by 36% in the case of Ru(bpy)_2dppz, 86% in the case of NDI-Ru_2 after the formation of double-stranded DNA. This result suggested that the metallointercalator was concentrated on the surface of gold electrode by its affinity to DNA duplex. In the case of NDI-Ru_2, we consider that electron was smoothly transferred from ruthenium to gold electrode through naphthalenediimide because NDI acts as an electron acceptor. In conclusion, we could discriminate between double stranded and single stranded DNA from the magnitude of the photocurrent using a ruthenium intercalator.

在这项研究中，我们使用光到电动转换开发了一种新型的DNA双链体检测系统。 DNA双链体形成的检测原理如下。 1）制备通过单链DNA修饰的金电极。 2）当形成双链DNA时，芳族复合物将插入并集中在电极表面上。 3）然后将可见光照射到电极上，将观察到由芳族复合物激发引起的光电流。我们使用携带DNA插入配体的polypyridyl ruthenium ruthenium复合物进行DNA检测。 RU（BPY）_3用作对照化合物。 RU（BPY）_2DPPZ用作带有DNA插入零件的钌配合物。此外，我们将ndi-ru_2合成为载有氟芬综合体的螺纹介导器。这种类型的化合物与DNA双链体具有较小的解离速率，因为它具有连接到两个笨重组的螺纹嵌入零件，将DNA-IMMOMMOMMOMIMBILIZED GOLD电极浸入溶液中，其中包含光到电动转换式嵌入器（RU（BPY）_3，_3，_3，_3，_3，_3，_3，_3，_3，_3，_3，） ru（bpy）_2Dppz，ndi-ru_2）。当将可见光照射到电极的表面时，在三级胺作为电子供体的存在下发生连续的光电流。在RU（BPY）_2DPPZ的情况下，光电流的量增加了36％，在形成双链DNA后NDI-RU_2的情况下为86％。该结果表明，金属区域量表通过与DNA双链体的亲和力集中在金电极表面上。在NDI-RU_2的情况下，我们认为电子通过萘将电子顺畅地从氟他二酰亚胺传递到金电极，因为NDI充当电子受体。总之，我们可以使用ruthenium Intercarator区分双链和单链DNA与光电流的大小。