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  • Title: Tight regulation and modulation via a C1-regulated promoter in Escherichia coli and Pseudomonas aeruginosa.
    Author: Schofield DA, Westwater C, Dolan JW, Norris JS, Schmidt MG.
    Journal: Curr Microbiol; 2002 Jun; 44(6):425-30. PubMed ID: 12000993.
    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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