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Related Concept Videos

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Inducible Operons: lac Operon

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The DNA replication, transcription, and translation processes are intricately coupled in bacteria, allowing efficient gene expression and rapid protein synthesis. While this physical and functional coordination is advantageous, it introduces challenges that bacteria overcome through specific regulatory mechanisms.Coupling of Replication, Transcription, and TranslationThe coupling of replication, transcription, and translation is a hallmark of bacterial gene expression. As the replisome unwinds...
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The organization of prokaryotic genes in their genome is notably different from that of eukaryotes. Prokaryotic genes are organized, such that the genes for proteins involved in the same biochemical process or function are located together in groups. This group of genes, along with their regulatory elements, are collectively known as an operon. The functional genes in an operon are transcribed together to give a single strand of mRNA known as polycistronic mRNA.
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Global regulatory systems in bacteria enable rapid and coordinated responses to environmental changes by integrating sensory inputs with gene expression, ensuring efficient adaptation to fluctuating conditions. Key global regulatory mechanisms include regulons, two-component systems, sigma factors, and secondary messengers.Regulons and Global RegulatorsA regulon is a collection of genes and operons controlled by a common global regulator. These regulators enable bacteria to prioritize resource...
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Quorum sensing is a mechanism of bacterial communication that enables coordinated gene expression in response to changes in population density. This facilitates collective behaviors that enhance survival, resource acquisition, and ecological adaptation. This process relies on small signaling molecules called autoinducers that accumulate as bacterial populations grow. When a critical threshold concentration of autoinducers is reached, bacterial cells collectively modify gene expression,...
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Updated: Apr 24, 2026

Applying an Inducible Expression System to Study Interference of Bacterial Virulence Factors with Intracellular Signaling
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A multi-tetracycline responsive induction system for gene expression in Bacillus subtilis.

Keira Reich-Veillette1, Amrita E Rhoads1, Elizabeth A Libby1

  • 1Department of Bioengineering, Northeastern University, Boston, Massachusetts, USA.

Applied and Environmental Microbiology
|April 23, 2026
PubMed
Summary

Researchers developed a new tetracycline-inducible system for Bacillus subtilis, offering precise gene expression control. This robust system functions across various conditions, enhancing applications in biotechnology and microbiology.

Keywords:
Bacillus subtilisCRISPRianhydrotetracyclinechlortetracyclinedoxycyclinetetracycline-inducible system

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Tightly regulated gene expression systems are crucial for microbiology research and applications.
  • Bacillus subtilis, a GRAS organism, lacks robust, tunable inducible systems adaptable to diverse growth conditions.
  • Existing tetracycline systems require optimization for B. subtilis's specific needs.

Purpose of the Study:

  • To engineer a sensitive, tightly regulated, and homogeneous tetracycline-inducible gene expression system for Bacillus subtilis.
  • To ensure the system's robustness across various media and growth conditions.
  • To provide a versatile tool for precise gene expression titration in B. subtilis.

Main Methods:

  • Engineered a codon-harmonized tetR repressor specific to B. subtilis.
  • Developed enhanced promoter and operator architecture for improved regulation.
  • Utilized a modular shuttle vector for stable, single-copy chromosomal integration.
  • Tested system performance with anhydrotetracycline, chlortetracycline, and doxycycline.

Main Results:

  • Achieved approximately 1,000-fold dynamic range with minimal leaky expression.
  • Demonstrated effective induction using subinhibitory concentrations of multiple tetracycline derivatives.
  • System showed robustness in rich media, minimal media, and on colonies.
  • Successfully applied the system to titrate sgRNA expression for CRISPRi applications.

Conclusions:

  • Developed a well-characterized, versatile tetracycline-inducible system for B. subtilis.
  • The system offers sensitive, tightly regulated, and homogeneous gene expression.
  • Enables precision titration for diverse applications including physiology studies, pathway optimization, and biotechnology.
  • Demonstrated compatibility with other genetic control systems like CRISPRi.