Amperometric DNA sensor using the pyrroquinoline quinone glucose dehydrogenase-avidin conjugate

被引：35

作者：

Ikebukuro, K ^{[1
]}

Kohiki, Y ^{[1
]}

Sode, K ^{[1
]}

机构：

[1] Tokyo Univ Agr & Technol, Fac Technol, Dept Biotechnol, Koganei, Tokyo 1848588, Japan

来源：

BIOSENSORS & BIOELECTRONICS | 2002年 / 17卷 / 11-12期

关键词：

pyrroquinoline quinone glucose dehydrogenase ((PQQ)GDH); DNA sensor; amperometry; (PQQ)GDH-avidin conjugate; Salmonella virulence invA gene; carbon paste electrode;

D O I：

10.1016/S0956-5663(02)00101-X

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

A new amperometric DNA sensor was constructed using a pyrroquinoline quinone glucose dehydrogenase ((PQQ)GDH) conjugated with avidin. Our aim was to specifically detect the DNA sequence of the invA virulence gene from the pathogenic bacterium Salmonella. Probe DNA with a sequence complementary to that of a specific fragment of the invA gene was immobilized onto a carbon paste electrode. After hybridization with biotinylated target DNA, (PQQ)GDH-avidin conjugate was added and the resulting electric current was measured. The electric current is generated from glucose oxidization catalyzed by (PQQ)GDH via 1-methoxyphenazine methosulphate (m-PMS) electron mediator. The sensor response increased with the addition of glucose and in the presence of 6.3 mM glucose the response increased with increasing DNA in the range 5.0 x 10(-8) -1.0 x 10(-5) M. (C) 2002 Elsevier Science B.V. All rights reserved.

引用

页码：1075 / 1080

页数：6

共 25 条

[1]

ALTSCHUL SF, 1990, J MOL BIOL, V215, P403, DOI 10.1006/jmbi.1990.9999

[2] Hybridization assay at a disposable electrochemical biosensor for the attomole detection of amplified human cytomegalovirus DNA [J].

Azek, F ;

Grossiord, C ;

Joannes, M ;

Limoges, B ;

Brossier, P .

ANALYTICAL BIOCHEMISTRY, 2000, 284 (01) :107-113

[3] Mutation detection by electrocatalysis at DNA-modified electrodes [J].

Boon, EM ;

Ceres, DM ;

Drummond, TG ;

Hill, MG ;

Barton, JK .

NATURE BIOTECHNOLOGY, 2000, 18 (10) :1096-1100

[4] Reactive electrophoretic activation of a microelectrode for enzyme-amplified recognition and for melting temperature determination of 105 copies of a simple oligonucleotide [J].

de Lumley-Woodyear, T ;

Caruana, DJ ;

Campbell, CN ;

Heller, A .

ANALYTICAL CHEMISTRY, 1999, 71 (02) :394-398

[5]

GALAN JE, 1992, J BACTERIOL, V174, P4338

[6] Electrochemical study of electron transport through thin DNA films [J].

Hartwich, G ;

Caruana, DJ ;

de Lumley-Woodyear, T ;

Wu, YB ;

Campbell, CN ;

Heller, A .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1999, 121 (46) :10803-10812

[7] Microfabricated disposable DNA sensor for detection of hepatitis B virus DNA [J].

Hashimoto, K ;

Ito, K ;

Ishimori, Y .

SENSORS AND ACTUATORS B-CHEMICAL, 1998, 46 (03) :220-225

[8] Construction and characterization of mutant water-soluble PQQ glucose dehydrogenases with altered Km values -: Site-directed mutagenesis studies on the putative active site [J].

Igarashi, S ;

Ohtera, T ;

Yoshida, H ;

Witarto, AB ;

Sode, K .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1999, 264 (03) :820-824

[9] Ferrocene-oligonucleotide conjugates for electrochemical probing of DNA [J].

Ihara, T ;

Maruo, Y ;

Takenaka, S ;

Takagi, M .

NUCLEIC ACIDS RESEARCH, 1996, 24 (21) :4273-4280

[10]

Karp M, 1996, BIOTECHNIQUES, V20, P452

← 1 2 3 →