Abstract
We describe the development and analysis of a novel promoter system regulated by the bacteriophage P1 temperature-sensitive C1 repressor. Using transcriptional fusions to the lacZ reporter gene to monitor gene expression, we show that the ratio of induction/repression can be up to 1500-fold in Escherichia coli. The promoters exhibited extremely tight repression and could be modulated over a range of temperatures. The utility of the promoter system was tested in Pseudomonas aeruginosa. C1 effectively repressed transcription; however, only modest induction was achieved. To increase the levels of induction, the amount of c1 was modulated at the mRNA level by using a LacI-regulated promoter. This resulted in a 59-fold induction in gene expression under inducing conditions. As the promoter system was constructed in a broad-host range vector and utilized the C1 repressor from a broad-host range phage, the system will provide the potential for controlled gene expression in Gram-negative bacteria. >
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Received: 19 July 2001 / Accepted: 25 September 2001
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Schofield, D., Westwater, C., Dolan, J. et al. Tight Regulation and Modulation via a C1-Regulated Promoter in Escherichia coli and Pseudomonas aeruginosa . Curr Microbiol 44, 425–430 (2002). https://doi.org/10.1007/s00284-001-0015-7
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DOI: https://doi.org/10.1007/s00284-001-0015-7