Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Green Fluorescent Protein as a Marker for Monitoring a Pentachlorophenol Degrader Sphingomonas chlorophenolica ATCC39723

  • Oh, Eun-Taex (Department of Biological Engineering, Center for Advanced Bioseparation Technology, Inha University) ;
  • So, Jae-Seong (Department of Biological Engineering, Center for Advanced Bioseparation Technology, Inha University) ;
  • Kim, Byung-Hyuk (Division of Civil and Environmental Systems Engineering, Korea Maritime University) ;
  • Kim, Jong-Sul (Division of Life Science, University of Ulsan) ;
  • Koh, Sung-Cheol (Division of Civil and Environmental Systems Engineering, Korea Maritime University)
  • Published : 2004.09.01
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Abstract

Sphingomonas chlorophenolica ATCC39723 was successfully labeled with the gfp (green fluorescent protein) gene inserted into the pcpB gene by homologous recombination. As the gfp recombinant was easily distinguished from other indigenous organisms, the population of gfp recombinant was monitored after being released into the soil microcosms. Their population density dropped from 10$\^$8/ to 10$\^$6/ (cfu/$m\ell$) in the non-sterilized soil microcosms during the first 6 days. Moreover, the gfp recombinant was not detected even at lower dilution rates after a certain time period. The recombinant, however, survived for at least 28 days in the sterilized soil microcosms. Although the gfp recombinant did not degrade pentachlorophenol (PCP), this experiment showed the possibility of using gfp as a monitoring reporter system for S. chlorophenolica ATCC39723 and potentially other species of Sphingomonas.

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References

  1. Andersen, J.B., C. Sternberg, L.K. Poulsen, S.P. Bjorn, M. Givskov, and S. Molin. 1998. New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria. Appl. Environ. Microbiol. 64, 2240-2246
  2. Blackburn, J.W. and R.J. Hafker. 1993. The impact of biochemistry, bioavailability and bioactivity on the selection of bioremediation techniques. Trends Biotechnol. 11, 328-333
  3. Bloemberg, G.V., G.A. OTool, B.J.J. Lugtenberg, and T. Kolter. 1997. Green fluorescent protein as a marker for Pseudomonas spp. Appl. Environ. Microbiol. 63, 4543-4551
  4. Cai, M. and L. Xun. 2002. Organization and regulation pentachlorophenol-degrading genes in Sphingobium chlorophenolicum ATCC39723. J. Bacteriol. 184, 4672-4680
  5. Chalfie, M., Y. Tu, G. Euskirchen, W.W. Ward, and D.C. Prasher. 1994. Green fluorescent protein as a marker for gene expression. Science. 263, 802-805
  6. Chanama, S. and R.L. Crawford. 1997. Mutational analysis of pcpA and its role in pentachlorophenol degradation by Sphingomonas (Flavobacterium) chlorophenolica ATCC 39723. Appl. Environ. Microbiol. 63, 4833-4838
  7. Cubitt, A.B., R. Heim, S.R. Adams, A.E. Boyd, L.A. Gross, and R.Y. Tsien. 1995. Understanding, improving and using green fluorescent proteins. Trends in Biochem. Sci. 20, 448-455
  8. Edgehill, R.U. and R.K. Finn. 1982. Isolation, characterization and growth kinetics of bacteria metabolizing pentachlorophenol. European J. Appl. Microbiol. Biotechnol. 16, 179-184