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Host-vector system for phenol-degrading Rhodococcus erythropolis based on Corynebacterium plasmids

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Abstract

The strain Rhodococcus erythropolis CCM2595, which was shown to degrade phenol, was chosen for genetic studies. To facilitate strain improvement using the methods of gene manipulation, the technique of genetic transfer was introduced and cloning vectors were constructed. Using the plasmid pFAJ2574, an electrotransformation procedure yielding up to 7×104 transformants/μg DNA was optimized. Escherichia coli-R. erythropolis shuttle vectors were constructed using the replicons pSR1 and pGA1 from Corynebacterium glutamicum. The small vector pSRK21 (5.8 kb) provides six unique cloning sites and selection of recombinant clones using α-complementation of β-galactosidase in E. coli. This vector, exhibiting high segregational stability under non-selective conditions in R. erythropolis CCM2595, was applied to cloning and efficient expression of the gene coding for green fluorescent protein (gfpuv).

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Acknowledgements

We thank R. De Mot for the gift of pFAJ2574 and R. Kalscheuer and A. Steinbüchel for the gift of pNC9503. This work was supported by grant 526/01/0177 from the Grant Agency of the Czech Republic and by Institutional Research Concept no. AV0Z5020903.

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Authors and Affiliations

  1. Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220, Prague 4, Czech Republic

    M. Veselý, M. Pátek & J. Nešvera

  2. Department of Fermentation Chemistry and Bioengineering, Institute of Chemical Technology, 16628, Prague 6, Czech Republic

    A. Čejková, J. Masák & V. Jirků

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  1. M. Veselý
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  2. M. Pátek
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  3. J. Nešvera
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  4. A. Čejková
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  5. J. Masák
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  6. V. Jirků
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Correspondence to M. Pátek.

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Veselý, M., Pátek, M., Nešvera, J. et al. Host-vector system for phenol-degrading Rhodococcus erythropolis based on Corynebacterium plasmids. Appl Microbiol Biotechnol 61, 523–527 (2003). https://doi.org/10.1007/s00253-003-1230-x

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  • DOI: https://doi.org/10.1007/s00253-003-1230-x

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