Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Iron Chelator-Inducible Expression System for Escherichia coli

  • Lim, Jae-Myung (Omics and Integration Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Hong, Mi-Ju (Omics and Integration Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Seong-Hun (Omics and Integration Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Oh, Doo-Byoung (Omics and Integration Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kang, Hyun-Ah (Department of Life Science, College of Natural Science, Chung-Ang University) ;
  • Kwon, Oh-Suk (Omics and Integration Research Center, Korea Research Institute of Bioscience and Biotechnology)
  • Published : 2008.08.31
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Abstract

The $P_{entC}$ promoter of the entCERA operon encoding enzymes for enterobactin biosynthesis in Escherichia coli is tightly regulated by the availability of iron in the culture medium. In iron-rich conditions, the $P_{entC}$ promoter activity is strongly repressed by the global transcription regulator Fur (ferric uptake regulator), which complexes with ferrous ions and binds to the Fur box 19-bp inverted repeat. In this study, we have constructed the expression vector pOS2 containing the $P_{entC}$ promoter and characterized its repression, induction, and modulation by quantifying the expression of the lacZ reporter gene encoding $\beta$-galactosidase. $\beta$-Galactosidase activities of E. coli transformants harboring pOS2-lacZ were highly induced in the presence of divalent metal ion chelators such as 2,2'-dipyridyl and EDTA, and were strongly repressed in the presence of excess iron. It was also shown that the basal level $\beta$-galactosidase expression by the $P_{entC}$ promoter was drastically decreased by incorporating the fur gene into the expression vector. Since the newly developed iron chelator-inducible expression system is efficient and cost-effective, it has wide applications in recombinant protein production.

Keywords

References

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