Iron chelator-inducible expression system for Escherichia coli
- Authors
- Lim, Jae-Myung; Hong, Mi-Ju; Kim, Seonghun; Oh, Doo-Byoung; Kang, Hyun Ah; Kwon, Ohsuk
- Issue Date
- Aug-2008
- Publisher
- KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
- Keywords
- E. coli; iron chelator; Fur; expression system
- Citation
- JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, v.18, no.8, pp 1357 - 1363
- Pages
- 7
- Journal Title
- JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
- Volume
- 18
- Number
- 8
- Start Page
- 1357
- End Page
- 1363
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/23666
- ISSN
- 1017-7825
1738-8872
- Abstract
- The P-entC promoter of the entCEBA 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 P-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-inclucible expression system is efficient and cost-effective, it has wide applications in recombinant protein production.
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