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

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Wattiau, Pierre [UCL] Springael, Dirk [UCL] Agathos, Spiros N. [UCL] Wuertz, S

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.

Document type Article de périodique (Journal article) – Journal Article, Research Support, Non-U.S. Gov't
Access type Accès restreint
Publication date 2002
Journal information "Applied microbiology and biotechnology" - Vol. 59, no. 6, p. 700-5 (2002)
Peer reviewed yes
issn 0175-7598
Publication status Publié
Affiliation UCL - AGRO/CABI - Département de chimie appliquée et des bio-industries
MESH Subject DNA, Bacterial - chemistry - genetics ; Electroporation ; Escherichia coli - genetics ; Genetic Markers - genetics ; Green Fluorescent Proteins ; Luminescent Proteins - chemistry ; Microscopy, Confocal ; Mycobacterium - genetics - metabolism ; Plasmids - genetics ; Polycyclic Hydrocarbons, Aromatic - metabolism ; Promoter Regions, Genetic ; Recombinant Proteins - chemistry - genetics ; Sequence Analysis, DNA ; Transformation, Bacterial - genetics
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Bibliographic reference Wattiau, Pierre ; Springael, Dirk ; Agathos, Spiros N. ; Wuertz, S. Use of the pAL5000 replicon in PAH-degrading mycobacteria: application for strain labelling and promoter probing.. In: Applied microbiology and biotechnology, Vol. 59, no. 6, p. 700-5 (2002)
Permanent URL http://hdl.handle.net/2078.1/24012