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KMID : 0624620100430070468
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BMB Reports
2010 Volume.43 No. 7 p.468 ~ p.473
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Detection of the cell wall-affecting antibiotics at sublethal concentrations using a reporter Staphylococcus aureus harboring drp35 promoter - lacZ transcriptional fusion
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Mondal Rajkrishna
Chanda Palas K
Bandhu Amitava
Jana Biswanath
Sau Subrata
Lee Chia-Y
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Abstract
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Previously, various inhibitors of cell wall synthesis induced the drp35 gene of Staphylococcus aureus efficiently. To determine whether drp35 could be exploited in antistaphylococcal drug discovery, we cloned the promoter of drp35 (Pd) and developed different biological assay systems using an engineered S. aureus strain that harbors a chromosomally-integrated Pd - lacZ transcriptional fusion. An agarose-based assay showed that Pd is induced not only by the cell wall-affecting antibiotics but also by rifampicin and ciprofloxacin. In contrast, a liquid medium-based assay revealed the induction of Pd specifically by the cell wall-affecting antibiotics. Induction of Pd by sublethal concentrations of cell wall-affecting antibiotics was even assessable in a microtiter plate assay format, indicating that this assay system could be potentially used for high-throughput screening of new cell wall-inhibiting compounds.
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KEYWORD
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Agarose-based assay and microtiter plate assay, Cell wall-affecting antibiotics, drp35, Staphylococcus aureus
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