Development of enzyme free sensitive gene detection system based ontemplate-directed formation of luminescent lanthanide complexe

我们提出了一种简单的比色SNP（单核苷酸多态性）分析技术，通过DNA模板的发光镧系离子（Ln（III）：Tb（III）或Eu（III））和两个ODN（寡脱氧核糖核苷酸）共轭carrying金属螯合剂之间的合作络合。EDTA（乙二胺四乙酸）和1，10-菲咯啉（phen）共价连接到ODNs上，形成偶联物，作为捕获和敏化探针。偶联物的序列被设计为与靶形成三元串联双链体，其中它们的双链体单元彼此面对，提供容纳Ln（III）的微环境。在完全匹配的靶DNA存在下，可以清楚地观察到Tb（III）和Eu（III）离子的特征发射。另一方面，含有一个碱基置换的突变体的混合溶液的排放量是微不足道的。

We present the facile technique of colorimetric SNP (single nucleotide polymorphism)analysis through DNA-templated cooperative complexation between a luminescent lanthanideion (Ln(III): Tb(III) or Eu(III)) and two ODN (oligodeoxyribonucleotide) conjugatescarrying a metal chelator. EDTA (ethylenediaminetetraacetic acid)) and1,10-phenanthroline (phen) were covalently attached to ODNs to form conjugates forapplication as capture and sensitizer probes. The sequences of the conjugates weredesigned so as to form a ternary tandem duplex with the target, where their auxiliaryunits face each other, providing a microenvironment to accommodate Ln(III). Thecharacteristic emissions of the Tb(III) and Eu(III) ions were clearly observed in thepresence of the full-matched target DNA. On the other hand, the emissions from the mixedsolutions with the mutant containing one base displacement were marginal.