Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Coordination of Gene Expression Processes in Bacteria01:29

Coordination of Gene Expression Processes in Bacteria

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...
Gene Regulation in Microbial Communities: Quorum Sensing01:28

Gene Regulation in Microbial Communities: Quorum Sensing

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,...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Targeted genomic editing of human gut Bacteroides species based on CRISPR-associated transposases.

Cell systems·2026
Same author

Author Correction: A clinic-responder-derived defined microbial consortium enhances anti-PD-1 immunotherapy efficacy in mice.

Nature microbiology·2026
Same author

Salmonella-derived haem inhibits macrophage phagocytosis and promotes infection in mice.

Nature microbiology·2026
Same author

A clinic-responder-derived defined microbial consortium enhances anti-PD-1 immunotherapy efficacy in mice.

Nature microbiology·2026
Same author

Global genomic insights into the antimicrobial resistance of <i>Escherichia marmotae</i>.

Zoological research·2025
Same author

Development of NAFLD-Specific Human Liver Organoid Models on a Microengineered Array Chip for Semaglutide Efficacy Evaluation.

Cell proliferation·2025

Related Experiment Video

Updated: Jun 29, 2026

Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems
06:18

Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems

Published on: April 26, 2019

A novel differential expression system for gene modulation in Mycobacteria.

Xiao-Yong Fan1, Hui Ma, Jian Guo

  • 1Department of Microbiology, School of Life Sciences, Fudan University, Shanghai, China. xyfan008@hotmail.com

Plasmid
|October 7, 2008
PubMed
Summary

Researchers optimized the Mycobacterium tuberculosis furA promoter (pfurA) for genetic manipulation. Mutants achieved a tenfold increase in activity, enabling differential antigen expression for tuberculosis vaccine development.

More Related Videos

Live Cell Fluorescence Microscopy to Observe Essential Processes During Microbial Cell Growth
07:28

Live Cell Fluorescence Microscopy to Observe Essential Processes During Microbial Cell Growth

Published on: November 24, 2017

Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX
10:09

Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX

Published on: June 27, 2017

Related Experiment Videos

Last Updated: Jun 29, 2026

Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems
06:18

Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems

Published on: April 26, 2019

Live Cell Fluorescence Microscopy to Observe Essential Processes During Microbial Cell Growth
07:28

Live Cell Fluorescence Microscopy to Observe Essential Processes During Microbial Cell Growth

Published on: November 24, 2017

Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX
10:09

Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX

Published on: June 27, 2017

Area of Science:

  • Microbiology
  • Molecular Biology
  • Immunology

Background:

  • Tuberculosis (TB) is a significant global health challenge.
  • Genetic manipulation of Mycobacterium tuberculosis (M.tb) is vital for understanding pathogenesis and developing new TB treatments.
  • Optimizing promoter activity is key for effective genetic tools in mycobacteria.

Purpose of the Study:

  • To generate and characterize mutants of the M.tb furA gene operator/promoter (pfurA) to enhance promoter activity.
  • To establish a system for differential expression of antigens in mycobacterial strains.
  • To explore the potential for developing recombinant BCG vaccine candidates.

Main Methods:

  • Generation of pfurA mutants with specific codon changes and substitutions in the FurA binding region.
  • Quantification of promoter activity using a lacZ gene-fusion reporter system.
  • In vivo expression analysis in Mycobacterium bovis BCG during intracellular growth and macrophage infection.
  • Expression of M.tb chimeric antigen Ag856A2 in recombinant Mycobacterium smegmatis and BCG strains.

Main Results:

  • A GTG to ATG codon change doubled beta-galactosidase activity.
  • A 6-bp substitution in the AT-rich region increased activity 4-6 fold.
  • Combining both mutations resulted in a tenfold increase in activity compared to the prototype pfurA.
  • pfurA variants showed continuous in vivo expression and early induction upon macrophage infection.
  • Differential expression of antigen Ag856A2 was achieved in recombinant strains.

Conclusions:

  • Optimized pfurA mutants provide significantly enhanced promoter activity for mycobacterial genetic manipulation.
  • The developed differential expression system is versatile for studying gene regulation and expressing target antigens.
  • This system facilitates the development of novel recombinant BCG vaccine candidates for TB and other infectious diseases.