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Related Concept Videos

Reporter Genes02:11

Reporter Genes

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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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Related Experiment Video

Updated: Apr 5, 2026

Applying an Inducible Expression System to Study Interference of Bacterial Virulence Factors with Intracellular Signaling
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An IPTG Inducible Conditional Expression System for Mycobacteria.

Sudha Ravishankar1, Anisha Ambady1, Haripriya Ramu1

  • 1AstraZeneca India Pvt Ltd, Bellary Road, Hebbal, Bengaluru, Karnataka, India.

Plos One
|August 7, 2015
PubMed
Summary
This summary is machine-generated.

We developed new non-replicating isopropylthiogalactoside (IPTG) inducible vectors for mycobacteria. These vectors enable precise control of gene expression, crucial for drug discovery and target validation in bacterial pathogens.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Drug Discovery

Background:

  • Conditional expression systems are vital for studying gene essentiality and target vulnerability in drug discovery.
  • Mycobacterial research benefits from precise gene regulation tools to mimic knockout conditions and assess target depletion effects.
  • Existing isopropylthiogalactoside (IPTG) inducible systems in mycobacteria often utilize replicating plasmids, limiting versatility.

Purpose of the Study:

  • To develop and characterize a versatile set of non-replicating IPTG-inducible vectors for mycobacteria.
  • To evaluate the regulatory efficiency of single versus double lac operator systems for tight gene expression control.
  • To demonstrate the utility of the developed system for bacterial target validation and inhibitor screening.

Main Methods:

  • Construction of non-replicating IPTG-inducible vectors with single and double lac operators.
  • Monitoring of β-galactosidase expression in Mycobacterium smegmatis to assess gene regulation.
  • Generation of conditional expression strains (inhA, rpoB, ftsZ) via homologous recombination.
  • Analysis of growth kinetics of conditional strains across varying IPTG concentrations.
  • Screening of small molecule libraries using conditional strains to identify target-specific inhibitors.

Main Results:

  • A double lac operator vector demonstrated significantly tighter gene expression control with no detectable leaky expression compared to a single lac operator vector.
  • Conditional expression strains for inhA, rpoB, and ftsZ showed IPTG-dependent growth modulation, confirming effective target regulation.
  • The system successfully identified target-specific inhibitors when screening focused libraries against inhA and rpoB conditional strains.

Conclusions:

  • The developed non-replicating IPTG-inducible vectors provide a robust and versatile tool for generating conditional expression strains in mycobacteria.
  • The double lac operator system offers superior control over gene expression, essential for accurate target validation and drug discovery.
  • This inducible system facilitates the identification of novel drug targets and the screening of potential inhibitors in mycobacterial research.