Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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In Situ Monitoring of Biofilm Formations of Escherichia coli and Pseudomonas putida by Use of Lux and GFP Reporters

  • Khang, Youn-Ho (Department of Applied Microbiology, Yeungnam University) ;
  • Rober S. Burlage (Environmental Science Division, Oak Ridge National Laboratory)
  • Published : 1998.06.01
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Abstract

A plasmid vector containing two reporter genes, mer-lux and lac-GFP, was transformed to both Escherichia coli and Pseudomonas putida. Their cellular activities and biofilm characteristics were investigated in flow-cell units by measuring bioluminescent lights and fluorescent levels of GFP. Bioluminescence was effective to monitor temporal cell activities, whereas fluorescent level of GFP was useful to indicate the overall cell activities during biofilm development. The light production rates of E. coli and P. putida cultures were dependent upon concentrations of HgCl2. Mercury molecules entrapped in P. putida biofilms were hardly washed out in comparison with those in E. coli biofilms, indicating that P. putida biofilms may have higher affinity to mercury molecules than E. coli biofilms. It was observed that P. putida expressed GFP cDNA in biofilms but not in liquid cultures. This may indicate that the genetic mechanisms of P. putida were favorably altered in biofilm conditions to make a foreign gene expression possible.

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