2'-anthraquinone-conjugated oligonucleotide as an electrochemical probe for DNA mismatch.

2'-anthraquinone-conjugated oligonucleotide as an electrochemical probe for DNA mismatch. Research Abstract Details 

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2'-anthraquinone-conjugated oligonucleotide as an electrochemical probe for DNA mismatch. Abstract Text:

Satoshi Kumamoto,Mariko Watanabe,Naoko Kawakami,Mitsunobu Nakamura,Kazushige Yamana,Satoshi Kumamoto,Mariko Watanabe,Naoko Kawakami,Mitsunobu Nakamura,Kazushige Yamana,Satoshi Kumamoto,Mariko Watanabe,Naoko Kawakami,Mitsunobu Nakamura,Kazushige Yamana,

We previously prepared the oligonucleotides (ODNs) conjugated to an anthraquinone (AQ) group via one carbon linker at the 2'-sugar position. When these modified ODNs bind to cDNA sequences, the AQ moiety can be intercalated into the predetermined base-pair pocket of a duplex DNA. In this paper, 2'-AQ-modified ODNs are shown to be an excellent electrochemical probe to clarify the effect of a mismatch base on the charge transfer (CT) though DNA. Two types of DNA-modified electrodes were constructed by assembly of disulfide-terminated 2'-AQ-ODN duplexes onto gold electrodes. One type of electrodes (system I) contains fully matched base pairs or a single-base mismatch in duplex DNA between the redox center and the electrode. The other (system II) consists of the mismatch but at the outside of the redox center. The modified electrodes were analyzed by cyclic voltammetry to estimate the CT rate through duplex DNA. In system I, the CT rate was found to be approximately 50 s (-1) for the fully matched AQ-ODN duplexes, while the CT rates of the mismatched DNA were considerably slower than that of the fully matched DNA. In system II, the AQ-ODN duplexes showed almost similar CT rates ( approximately 50 s (-1)) for the fully matched DNA and for the mismatched DNAs. The detection of a single-base mismatch was then performed by chronocoulometry (CC). All the DNA duplexes containing a mismatch base in system I gave the reduced electrochemical responses when compared to the fully matched DNA. In particular, the mismatched DNAs including G-A mismatch can be differentiated from fully matched DNA without using any electrochemical catalyst. We further tested the usefulness of single-stranded (ss) AQ-ODN immobilized on a gold electrode in the electrochemical detection of a single-base mismatch through hybridization assay. The ss-AQ-ODN electrodes were immersed in target-containing buffer at room temperature, and the CC measurements were carried out to see the changes in the integrated charge. Within 60 min, the mismatched DNA was clearly distinguishable by the CC differences from the fully matched target. Thus, the electrochemical hybridization assay provides an easy and convenient detection for DNA mutation that does not require any extra reagents, catalyst, target labeling, and washing steps.

2'-anthraquinone-conjugated oligonucleotide as an electrochemical probe for DNA mismatch. Publishing Authors By Initials

S Kumamoto,M Watanabe,N Kawakami,M Nakamura,K Yamana,S Kumamoto,M Watanabe,N Kawakami,M Nakamura,K Yamana,S Kumamoto,M Watanabe,N Kawakami,M Nakamura,K Yamana,

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2'-anthraquinone-conjugated oligonucleotide as an electrochemical probe for DNA mismatch. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Bioconjugate chemistry

VOLUME: 19

Page Numbers: 65-9

Journal Abbreviation: Bioconjug. Chem.

ISSN: 1043-1802

DAY: 8

MONTH: 11

YEAR: 2007

2'-anthraquinone-conjugated oligonucleotide as an electrochemical probe for DNA mismatch. Information

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LANGUAGE: eng

NlmUniqueID: 9010319

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Grant and Affiliation Information for 2'-anthraquinone-conjugated oligonucleotide as an electrochemical probe for DNA mismatch.

AFFILIATION: yamana@eng.u-hyogo.ac.jp.

Country: United States

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MEDLINETA: Bioconjug Chem

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