Development of a thermally regulated broad-spectrum promoter system for use in pathogenic gram-positive species.


:Selectively regulating gene expression is an essential molecular tool that is lacking for many pathogenic gram-positive bacteria. In this report, we describe the evaluation of a series of promoters regulated by the bacteriophage P1 temperature-sensitive C1 repressor in Enterococcus faecium, Enterococcus faecalis, and Staphylococcus aureus. Using the lacZ gene to monitor gene expression, we examined the strength, basal expression, and induced expression of synthetic promoters carrying C1 operator sites. The promoters exhibited extremely low basal expression and, under inducing conditions, gave high levels of expression (100- to 1,000-fold induction). We demonstrate that the promoter system could be modulated by temperature and showed rapid induction and that the mechanism of regulation occurred at the level of transcription. Controlled expression with the same constructs was also demonstrated in the gram-negative bacterium Escherichia coli. However, low basal expression and the ability to achieve derepression were dependent on both the number of mismatches in the C1 operator sites and the promoter driving c1 expression. Since the promoters were designed to contain conserved promoter elements from gram-positive species and were constructed in a broad-host-range plasmid, this system will provide a new opportunity for controlled gene expression in a variety of gram-positive bacteria.


Appl Environ Microbiol


Schofield DA,Westwater C,Hoel BD,Werner PA,Norris JS,Schmidt MG





Has Abstract


2003-06-01 00:00:00












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    pub_type: 杂志文章


    authors: Murinda SE,Roberts RF,Wilson RA

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    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Masuda S,Eda S,Ikeda S,Mitsui H,Minamisawa K

    更新日期:2010-04-01 00:00:00

  • Split-Root Assays Using Trifolium subterraneum Show that Rhizobium Infection Induces a Systemic Response That Can Inhibit Nodulation of Another Invasive Rhizobium Strain.

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    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Sargent L,Huang SZ,Rolfe BG,Djordjevic MA

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    abstract::[This corrects the article on p. 1239 in vol. 45.]. ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章



    更新日期:1983-07-01 00:00:00

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    abstract::Bacillus subtilis RB14, which showed antibiotic activities against several phytopathogens in vitro by producing the antibiotics iturin A and surfactin, was subjected to a pot test to investigate its ability to suppress damping-off of tomato seedlings caused by Rhizoctonia solani. To facilitate recovery from soil, B. s...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Asaka O,Shoda M

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    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


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    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


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    pub_type: 杂志文章


    authors: Sommaruga R,Hofer JS,Alonso-Sáez L,Gasol JM

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    pub_type: 杂志文章


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    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Marquet-Van der Mee N,Audurier A

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    journal_title:Applied and environmental microbiology

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    pub_type: 杂志文章


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    authors: Chakroun M,Ferré J

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    pub_type: 杂志文章


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    pub_type: 杂志文章


    authors: Lovley DR

    更新日期:1985-06-01 00:00:00

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    pub_type: 杂志文章


    authors: Takenaka S,Pitts B,Trivedi HM,Stewart PS

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    pub_type: 杂志文章


    authors: Bonnet C,Diarrassouba F,Brousseau R,Masson L,Topp E,Diarra MS

    更新日期:2009-11-01 00:00:00