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Use of the pAL5000 replicon in PAH-degrading mycobacteria: application for strain labelling and promoter probing

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Abstract.

Three environmental Mycobacterium strains (LB501T, LB307T and VM552) able to degrade anthracene, phenanthrene or pyrene, respectively, were successfully electroporated with pAL5000-based plasmids containing the green fluorescent protein (gfp) gene of Aequoria victoria under the control of the hsp60 promoter of Mycobacterium bovis following a slightly modified standard procedure. Transformants showed irregular gfp expression profiles. Four plasmid derivatives were constructed that contained gene promoters isolated from, and adapted to, gene expression in polycyclic aromatic hydrocarbon (PAH)-degrading mycobacteria. One derivative directed strong and homogeneous expression of GFP, allowing dual analysis of both GFP- and PAH-derived fluorescence as assessed by confocal laser scanning microscopy. The results reported here demonstrate the suitability of the pAL5000 replicon for the development of recombinant DNA-based studies in PAH-degrading Mycobacterium spp.

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Author notes

  1. D. Springael

    Present address: Laboratory of Soil Fertility and Soil Biology, Catholic University of Leuven, Kasteelpark Arenberg 20, 3001 Heverlee, Belgium, Belgium

  2. S. Wuertz

    Present address: Department of Civil and Environmental Engineering, University of California, Davis, One Shields Avenue, Davis, CA 95616, USA, USA

Authors and Affiliations

  1. Catholic University of Louvain, Bioengineering Unit, Place Croix du Sud 2, Box 19, 1348 Louvain-la-Neuve, Belgium, Belgium

    P. Wattiau & S. N. Agathos

  2. Vlaamse Instelling voor Technologische Onderzoek, Boeretang 200, 2400 Mol, Belgium, Belgium

    D. Springael

  3. Institute of Water Quality Control and Waste Management, Technical University of Munich, Am Coulombwall, 85748 Garching, Germany, Germany

    S. Wuertz

Authors
  1. P. Wattiau
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  2. D. Springael
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  3. S. N. Agathos
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  4. S. Wuertz
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Correspondence to P. Wattiau.

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Wattiau, P., Springael, D., Agathos, S.N. et al. Use of the pAL5000 replicon in PAH-degrading mycobacteria: application for strain labelling and promoter probing. Appl Microbiol Biotechnol 59, 700–705 (2002). https://doi.org/10.1007/s00253-002-1081-x

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  • DOI: https://doi.org/10.1007/s00253-002-1081-x

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