MOLECULAR WIRING TYPE DNA SENSOR

分子布线型 DNA 传感器

基本信息

批准号：
11650851
负责人：
KUBO Izumi
金额：
$ 1.47万
依托单位：
Soka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

Electrochemical detection of specific DNA sequence of human papiroma virus was investigated using the gold electrode immobilized with probe DNA through a thiol group and tris(1,10-phenanthroline) cobalt (II) complex [Co(phen)_3^<2+>] as a hybridization indicator.25-mer probe DNA modified with a thiol group at 5' end was immobilized on a gold electrode. Tris(1,10-phenanthroline) cobalt (II) chloride was prepared according to previously reported procedure. The gold electrode immobilized with probe DNA was immersed into buffer solution (pH 7.0) containing target DNA.Hybridization reaction was performed for an hour at 65 C。.After the hybridization, the electrode was immersed into Co(phen)_3^<2+>/0.1M phosphate buffer (pH 7.0). Cyclic voltammetry was performed in Co(phen)_3^<2+>/0.1 M phosphate buffer (pH 7.0).Anodic peak current of cyclic voltammograms of 0.1 mM Co(phen)_3^<2+>/0.1 M phosphate buffer (pH 7.0) using the probe DNA-modified gold electrode hybridized with target DNA was larger … More than that using the probe DNA-modified gold electrode.When the concentration of Co(phen)_3^<2+> was 0.1 mM, the response currents depended on target DNA concentration. However, at another Co(phen)_3^<2+> concentrations of 0.2 mM, 0.3mM and 1.0 mM examined, the response currents did not clearly depend on target DNA concentration.The response currents were linearly related to target DNA concentration in the range from 10^<-7> M to 10^<-4> M with a slope of 10.5 nA / decade. Similar measurement was performed using the probe DNA-modified gold electrode reacted with 25-mer poly-deoxy A as non-complementary DNA.The response currents were hardly increased in the same range. On the other hand, all currents were slightly decreased from that using probe DNA-modified electrode. It seems that the surface of DNA-modified electrode suffer from chemical alteration because the electrode was repeatedly used increasing the concentration of target DNA (or poly-deoxy A) after one measurement. From these results, it was shown that a certain sequence of DNA, which is complementary to probe DNA, can be detected using cobalt phenanthroline complex and probe DNA-modified gold electrode. Less

使用与探针DNA固定的金电极通过硫醇基和Tris（1,10-磷酸氨基氨基氨酸）钴（II）复合物[CO（PEN）_3^<2+>]固定在探针DNA的特定DNA序列的电化学检测中，以杂交指标为25-Mer prog impior.25-摩布imob impiel a Impiel at thiol thiol atiol atiol atiol attiol attiol atiol atiol atiol atiol atiol atiol at'thiol thiol组。根据先前报道的程序，制备了Tris（1,10-磷酸）钴（II）氯化物。将固定在探针DNA的金电极浸入含有靶DNA的缓冲溶液（pH 7.0）中。在65 C时进行一个小时的杂交反应。杂交后，将电极浸入CO（PEN）_3^^<2+>/0.1M/0.1M磷酸盐燃料（pH 7.0）中。 Cyclic voltammetry was performed in Co(phen)_3^<2+>/0.1 M phosphate buffer (pH 7.0).Anodic peak current of cyclic voltammograms of 0.1 mM Co(phen)_3^<2+>/0.1 M phosphate buffer (pH 7.0) using the probe DNA-modified gold electrode hybridized with target DNA was larger … More than That using the probe DNA修饰的金电极。当CO（PEN）_3^<2+>的浓度为0.1 mm时，响应电流取决于靶DNA浓度。但是，在另一个CO（PEN）_3^<2+>浓度为0.2 mm，0.3 mm和1.0 mM的浓度下，响应电流并不明确取决于靶DNA浓度。响应电流与10^<-7> M至10^<-4> M s slope的靶向DNA浓度线性相关。使用探针DNA修饰的金电极与25-mer聚脱氧A反应为非平均DNA进行了类似的测量。在同一范围内，响应电流几乎不会增加。另一方面，使用探针DNA修饰的电极略微降低了所有电流。看来DNA修饰的电极的表面会遭受化学改变的影响，因为该电极被反复使用，在一次测量后增加了靶DNA（或聚脱氧A）的浓度。从这些结果中可以证明，可以使用钴酚类复合物和探针DNA修饰的金电极检测到某种与探针DNA互补的DNA序列。较少的