| Project/Area Number |
18550156
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Chemistry related to living body
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| Research Institution | University of Hyogo |
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Principal Investigator |
YAMANA Kazushige University of Hyogo, Materials Science ancl Chemistry, Professor (70192408)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAMURA Mitsunobu University of Hyogo, Materials Science and Chemistry, Associate Professor (50285342)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥4,190,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥390,000)
Fiscal Year 2007: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2006: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | (1)Redox-modified DNA / (2)Self-assembly / (3)Electrochemistry / (4)SNP detection / 電子移動 / 鎖交換 / 電気化学 / DNAチップ |
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| Research Abstract |
Redox-resposive DNA assembled monolayer at a metal surface is important in relation to developments of electrochemical DNA sensors and DNA-based electronic devices.
We previously prepared 2-O-(2-anthraquinoylmethyl)uridine-containing DNA fragments and found that they can bind selectively and strongly to their complementary DNA sequences. Importantly, the anthraquinone (AQ) moiety can be placed at the predetermined base-pair pocket in double helical DNA. We anticipated that the AQ-modified DNA could be used as an electrochemical probe for DNA sequence determination. The main purpose of the present research is to investigate electrochemical properties of the AQ-modified DNA assembled on a gold electrode surface.
First, we prepared AQ-modified DNA duplex attached via thiol-modification onto a gold electrode. One assembled DNA monolayer (system 1) has a single base mismatch between the redox-center and the electrode surface. And the other (system II) consists of the same mismatch but at the
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outside of the redox-center. Electrochemical measurements revealed. that the electron transfer (ET) rate for the fully matched DNA of system I was estimated to be ~50s^<-1>. The mismatch-containing DNA of system I indeed showed much slower ET rate. As expected, in system II, the ET rates for fully matched and mismatched DNA were similar (-50s^<-1>).
Then, the presence of mismatch in DNA was determined from the reduced electrochemical responses by a conventional measurement. The technique can be applied to hybridization assays of SNP typing. The important features are that the assay does not require any extra reagents, catalyst, target labeling, and washing steps. Therefore, the present method is a promising way for simple and convenient detection of single base mismatches in DNA.
We further investigated other possibility for SNP typing that is based on DNA strand exchange at a metal surface. Redox-modified double strand DNA was assembled as monolayer at a metal surface. We have obtained several experimental results that this type of redox-DNA bound electrodes may be used for SNP detection by a simple electrochemical assays. DNA strand exchange has been applied to developments of aptamer-based biosensors. The results on strand exchange approach will be reported in detail in the next grant period. Less
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