A protein-based oxygen biosensor for high-throughput monitoring of cell growth and cell viability

被引：20

作者：

Strianese, Maria ^{[1
]}

Zauner, Gerhild ^{[1
]}

Tepper, Armand W. J. W. ^{[1
]}

Bubacco, Luigi ^{[2
]}

Breukink, Eefjan ^{[3
]}

Aartsma, Thijs J. ^{[4
]}

Canters, Gerard W. ^{[1
]}

Tabares, Leandro C. ^{[1
]}

机构：

[1] Leiden Univ, Leiden Inst Chem, NL-2300 RA Leiden, Netherlands

[2] Univ Padua, Dept Biol, I-30121 Padua, Italy

[3] Univ Utrecht, Dept Chem Biol & Organ Chem, NL-3584 CH Utrecht, Netherlands

[4] Leiden Univ, Leiden Inst Phys, NL-2300 RA Leiden, Netherlands

来源：

ANALYTICAL BIOCHEMISTRY | 2009年 / 385卷 / 02期

关键词：

Biosensors; Oxygen sensing; Cell viability; Antibiotic screening; FRET; PSEUDOMONAS-AERUGINOSA; ESCHERICHIA-COLI; COPPER PROTEINS; FLUORESCENCE; PEPTIDE; COMPLEX; ASSAYS; ACID; GENE;

D O I：

10.1016/j.ab.2008.11.017

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Fluorescently labeled hemocyanin has been previously proposed as an oxygen sensor. In this study, we explored the efficacy of this biosensor for monitoring the biological oxygen consumption of bacteria and its use in testing bacterial cell growth and viability of Escherichia coli, Pseudomonas aeruginosa, Paracoccus denitrificans, and Staphylococcus simulans. Using a microwell plate, the time courses for the complete deoxygenation of samples with different initial concentrations of cells were obtained and the doubling times were extracted. The applicability of our fluorescence-based cell growth assay as an antibacterial drug screening method was also explored. The results provide a proof-of-principle for a simple, quantitative, and sensitive method for high-throughput monitoring of prokaryotic cell growth and antibiotic susceptibility screening. (C) 2008 Elsevier Inc. All rights reserved.

引用

页码：242 / 248

页数：7

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