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Development of DNA Detection Method using Photo-induced Electron Transfer

开发利用光诱导电子转移的 DNA 检测方法

基本信息

批准号：
12555225
负责人：
IJIRO Kuniharu
金额：
$ 8.58万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12555225/
关键词：
Photo-induced electron transfer DNA monolayer nucleic acid base fluorescence life time gene high sensitivity non-label

项目摘要

Double-stranded DNA is expected to show function as a molecular wire based on the π-electron array structure, so-called "π-ways", of stacked nucleic acid bases. DNA-mediated photo-induced electron transfer between electron donors and acceptors which are intercalated or bound to base pairs in polynucleotides has been investigated. We have reported that the amphiphilic intercalator, C_<18>AO^+, monolayer can be intercalated into DNA and other polynucleotides at the air-water interface and the composite monolayer with ordered DNA array can be transferred on the solid support.Here we report, DNA-mediated photo-induced electron transfer in the monolayers containing electron donor and acceptor at the air-water interface. Binary mixed monolayers (an amphiphilic donor, C18AO+ and cationic amphiphile, 2C18N+2C1) and ternary mixture (an amphiphilic acceptor C12BphC5V2+) were prepared on the aqueous subphases of double-stranded DNA, RNA and other anionic polymers.Pressure-area isotherms and fluorescence spectra were measured simultaneously at the air-water interface by Langmuir film balance equipping fluorescence microscope. Electron transfer quenching of C18AO+ fluorescence by C12BphC5V2+ molecule are clearly. Fluorescence quenching was enhanced by DNA and RNA, but not affected by synthetic anionic polymers. This phenomenon is observed only in the double-stranded DNA and RNA. Therefore hybridization of single-stranded DNA and RNA with complementary DNA and RNA can be detected by the photo-induced electron transfer through DNA and RNA combined with the ternary monolayer.

双链DNA预期显示出作为基于堆叠的核酸碱基的π-电子阵列结构（所谓的“π-路”）的分子导线的功能。研究了DNA介导的光诱导电子转移，电子供体和受体插入或结合到多核苷酸中的碱基对上。本文报道了两亲性嵌入剂C_<18>AO^+在空气-水界面上可嵌入DNA及其它多核苷酸，并可在固体支持物上转移有序DNA阵列的复合单分子膜，在此我们报道了含电子给体和电子受体的单分子膜在空气-水界面上的DNA介导的光诱导电子转移。在双链DNA、RNA及其它阴离子聚合物水亚相上制备了二元混合单分子膜（两亲供体C18 AO+和阳离子两亲物2C 18 N +2C 1）和三元混合膜（两亲受体C12 BphC 5V 2+），用朗缪尔薄膜天平和荧光显微镜同时测定了气-水界面的压力-面积等温线和荧光光谱。C12 BphC 5V 2+分子对C18 AO+荧光的电子转移猝灭明显。荧光猝灭增强DNA和RNA，但不受合成阴离子聚合物。这种现象仅在双链DNA和RNA中观察到。因此，单链DNA和RNA与互补DNA和RNA的杂交可以通过与三元单层结合的DNA和RNA的光诱导电子转移来检测。