Electrochemical detection of pathogenic bacteria by using a glucose dehydrogenase fused zinc finger protein

We developed an electrochemical detection system for pathogenic bacteria by utilizing a glucose dehydrogenase-fused zinc finger protein (ZF-GDH), which could detect PCR products electrochemically without the need for DNA probe hybridization. Using ZF-GDH, we could specifically detect 10 copies of genomic DNA derived fromEscherichia coliO157.

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Development of an electrochemical detection system for measuring DNA methylation levels using methyl CpG-binding protein and glucose dehydrogenase-fused zinc finger protein

Biosensors and Bioelectronics ◽

10.1016/j.bios.2016.09.060 ◽

2017 ◽

Vol 93 ◽

pp. 118-123 ◽

Cited By ~ 12

Author(s):

Jinhee Lee ◽

Wataru Yoshida ◽

Koichi Abe ◽

Kazuhiko Nakabayashi ◽

Hironobu Wakeda ◽

...

Keyword(s):

Dna Methylation ◽

Electrochemical Detection ◽

Zinc Finger ◽

Zinc Finger Protein ◽

Detection System ◽

Binding Protein ◽

Glucose Dehydrogenase ◽

Finger Protein

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Zinc finger protein-based detection system of PCR products for pathogen diagnosis

Nucleic Acids Symposium Series ◽

10.1093/nass/nrn012 ◽

2008 ◽

Vol 52 (1) ◽

pp. 23-24 ◽

Cited By ~ 5

Author(s):

Y. Osawa ◽

H. Motoki ◽

T. Matsuo ◽

M. Horiuchi ◽

K. Sode ◽

...

Keyword(s):

Zinc Finger ◽

Zinc Finger Protein ◽

Detection System ◽

Finger Protein ◽

Pcr Products ◽

Pathogen Diagnosis

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Application of a Glucose Dehydrogenase-Fused with Zinc Finger Protein to Label DNA Aptamers for the Electrochemical Detection of VEGF

Sensors ◽

10.3390/s20143878 ◽

2020 ◽

Vol 20 (14) ◽

pp. 3878

Author(s):

Jinhee Lee ◽

Atsuro Tatsumi ◽

Kaori Tsukakoshi ◽

Ellie D. Wilson ◽

Koichi Abe ◽

...

Keyword(s):

Electrochemical Detection ◽

Zinc Finger ◽

Electrochemical Sensors ◽

Zinc Finger Protein ◽

Specific Binding ◽

Glucose Dehydrogenase ◽

Current Response ◽

Finger Protein ◽

Electrochemical Signal ◽

Endothelial Growth Factor

Aptamer-based electrochemical sensors have gained attention in the context of developing a diagnostic biomarker detection method because of their rapid response, miniaturization ability, stability, and design flexibility. In such detection systems, enzymes are often used as labels to amplify the electrochemical signal. We have focused on glucose dehydrogenase (GDH) as a labeling enzyme for electrochemical detection owing to its high enzymatic activity, availability, and well-established electrochemical principle and platform. However, it is difficult and laborious to obtain one to one labeling of a GDH-aptamer complex with conventional chemical conjugation methods. In this study, we used GDH that was genetically fused to a DNA binding protein, i.e., zinc finger protein (ZF). Fused GDH can be attached to an aptamer spontaneously and site specifically in a buffer by exploiting the sequence-specific binding ability of ZF. Using such a fusion protein, we labeled a vascular endothelial growth factor (VEGF)-binding aptamer with GDH and detected the target electrochemically. As a result, upon the addition of glucose, the GDH labeled on the aptamer generated an amperometric signal, and the current response increased dependent on the VEGF concentration. Eventually, the developed electrochemical sensor proved to detect VEGF levels as low as 105 pM, thereby successfully demonstrating the concept of using ZF-fused GDH to enzymatically label aptamers.

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Development of a Detection System for Epigenetic Modifications By Enzyme Fused Zinc Finger Protein

ECS Meeting Abstracts ◽

10.1149/ma2016-02/44/3269 ◽

2016 ◽

Keyword(s):

Zinc Finger ◽

Zinc Finger Protein ◽

Detection System ◽

Epigenetic Modifications ◽

Finger Protein

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Ultrasensitive Electrochemical Detection of miRNA-21 Using a Zinc Finger Protein Specific to DNA–RNA Hybrids

Analytical Chemistry ◽

10.1021/acs.analchem.6b04609 ◽

2017 ◽

Vol 89 (3) ◽

pp. 2024-2031 ◽

Cited By ~ 40

Author(s):

Chiew San Fang ◽

Kwang-sun Kim ◽

Byeongjun Yu ◽

Sangyong Jon ◽

Moon-Soo Kim ◽

...

Keyword(s):

Electrochemical Detection ◽

Zinc Finger ◽

Zinc Finger Protein ◽

Finger Protein

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Washing-Free Electrochemical Detection of Amplified Double-Stranded DNAs Using a Zinc Finger Protein

Analytical Chemistry ◽

10.1021/acs.analchem.8b00143 ◽

2018 ◽

Vol 90 (7) ◽

pp. 4776-4782 ◽

Cited By ~ 8

Author(s):

Chiew San Fang ◽

Kwang-sun Kim ◽

Dat Thinh Ha ◽

Moon-Soo Kim ◽

Haesik Yang

Keyword(s):

Electrochemical Detection ◽

Zinc Finger ◽

Zinc Finger Protein ◽

Finger Protein

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Detection of Pathogenic Bacteria by Using Zinc Finger Protein Fused with Firefly Luciferase

Analytical Chemistry ◽

10.1021/ac3018845 ◽

2012 ◽

Vol 84 (18) ◽

pp. 8028-8032 ◽

Cited By ~ 19

Author(s):

Koichi Abe ◽

Takenori Kumagai ◽

Chiaki Takahashi ◽

Aki Kezuka ◽

Yoshiyuki Murakami ◽

...

Keyword(s):

Zinc Finger ◽

Pathogenic Bacteria ◽

Zinc Finger Protein ◽

Firefly Luciferase ◽

Finger Protein

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Sensitive and direct electrochemical detection of double-stranded DNA utilizing alkaline phosphatase-labelled zinc finger proteins

The Analyst ◽

10.1039/c5an00623f ◽

2015 ◽

Vol 140 (12) ◽

pp. 3947-3952 ◽

Cited By ~ 13

Author(s):

Soodong Noh ◽

Dat Thinh Ha ◽

Haesik Yang ◽

Moon-Soo Kim

Keyword(s):

Alkaline Phosphatase ◽

Electrochemical Detection ◽

Zinc Finger ◽

Zinc Finger Protein ◽

Direct Detection ◽

Electrochemical Measurement ◽

Zinc Finger Proteins ◽

Double Stranded Dna ◽

Ito Electrode ◽

Finger Protein

We report sensitive and direct detection of specific bacterial dsDNA sequences using (i) the zinc finger protein labelled with alkaline phosphatase and (ii) electrochemical measurement at the ITO electrode.

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