Target-assembled tandem oligonucleotide systems based on exciplexes for detecting DNA mismatches and single nucleotide polymorphisms

被引：41

作者：

Bichenkova, EV ^{[1
]}

Savage, HE ^{[1
]}

Sardarian, AR ^{[1
]}

Douglas, KT ^{[1
]}

机构：

[1] Univ Manchester, Sch Pharm & Pharmaceut Sci, Wolfson Ctr Rat Struct Based Design Mol Diagnost, Manchester M13 9PL, Lancs, England

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2005年 / 332卷 / 04期

基金：

英国生物技术与生命科学研究理事会;

关键词：

exciplex; tandem oligonucleotide; DNA detection; fluorescence; solvent effects on DNA; single nucleotide polymorphism; self-assembled detector systems;

D O I：

10.1016/j.bbrc.2005.05.048

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We report the first exciplex-based split-probe system for DNA detection. The detector is split at a molecular level into signal-silent components which, before a signal is generated, must be assembled correctly into a particular three-dimensional arrangement. The model system comprises of two 8-mer oligonucleotides, complementary to neighbouring sites of a 16-mer DNA target, each equipped with moieties able to form an exciplex on correct, contiguous hybridization. The exciplex emits at similar to 480 nm with a large Stokes shift (135 nm). The extremely rigorous structural demands for exciplex formation and emission were achieved by careful structural design and by the discovery that high levels of certain organic solvents (especially trifluoroethanol) specifically favour emission of the DNA-mounted exciplex, probably the net result of the particular duplex structure and specific solvation of the exciplex partners. Inserts and mismatches can be effectively detected by this exciplex construct giving potential for single nucleotide polymorphism detection. (c) 2005 Elsevier Inc. All rights reserved.

引用

页码：956 / 964

页数：9