• Title/Summary/Keyword: drp35

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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

  • Mondal, Rajkrishna;Chanda, Palas K.;Bandhu, Amitava;Jana, Biswanath;Lee, Chia-Y.;Sau, Subrata
    • BMB Reports
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    • v.43 no.7
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    • pp.468-473
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    • 2010
  • 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 ($P_d$) and developed different biological assay systems using an engineered S. aureus strain that harbors a chromosomally-integrated $P_d$ - lacZ transcriptional fusion. An agarose-based assay showed that $P_d$ 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 $P_d$ specifically by the cell wall-affecting antibiotics. Induction of $P_d$ 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.

Isolation and Phylogenetic Characteristics of Exopolysaccharide Producing Bacteria in a Rhizosphere Soil of Medicinal Herbs (약초 근권토양 내 다당 생성세균 분리 및 계통학적 특성)

  • Lee, Hae-Ran;Kim, Ki-Kwhang;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.278-285
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    • 2010
  • We examined the distribution of exopolysaccharide (EPS) producing bacteria population in rhizosphere soils of domestic medicinal herbs; Angelica sinensis, Atractytodes japonica, Achyranthes japonica, Anemarrhena asphodeloides, and Astragalus membranaceus. Fifty-six percent of the total isolates from rhizosphere soil of Angelica sinensis were EPS producing bacteria, suggesting the dominance of EPS producing bacteria in rhizosphere soil of Angelica sinensis. EPS producing bacteria were enumerated in root system (rhizosphere soil, rhizoplane, inside of root) of Angelica sinensis. Bacterial density of rhizosphere soil, rhizoplane, and inside of root were distributed $9.0{\times}10^6CFU/g{\cdot}soil$, $7.0{\times}10^6CFU/g{\cdot}soil$, and $1.4{\times}10^3CFU/g{\cdot}soil$, respectively. EPS producing bacteria from rhizosphere soil were categorized into five major phylogenetic groups: Alphaproteobacteria (4 strains), Betaproteobacteria (6 strains), Firmicutes (2 strains), Actinobacteria (3 strains), and Bacteroidetes (1 strain) subdivisions. Also, the EPS producing isolates from rhizoplane were distributed as 7 strains in Alphaproteobacteria, 3 strains in Betaproteobacteria, 2 strains in Actinobacteria, 3 strains in Bacteroidetes, and 1 strain in Acidobacteria subdivisions. All of the EPS producing bacteria inside of root belong to genus Chitinophaga. Burkholderia caribiensis DR14, Terriglobus sp. DRP35, and Rhizobium hainanense SAP110 were selected in 112 EPS producing bacteria. These appeared to have produced high levels of exopolysaccharide 6,555 mpa.s, 3,275 mpa.s, and 1,873 mpa.s, respectively. The purified EPS was analyzed Bio-LC. As neutral sugars, glucose, galactose, mannose were detected and as amino sugars, galactosamine and glucosamine were detected. Especilally, analysis of Bio-LC showed that Rhizobium hainanense SAP110 produced glucose (60~89%) and glucosamine (8.5%) as major neutral sugar and amino sugar, respectively.