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

Repressible Operon: trp Operon01:21

Repressible Operon: trp Operon

2.2K
The trp operon in Escherichia coli exemplifies a repressible operon. It regulates the synthesis of tryptophan through repressor-mediated transcriptional control and attenuation. This dual regulatory mechanism ensures tryptophan biosynthesis occurs only when needed, conserving cellular resources.Structure of the trp OperonThe trp operon consists of five structural genes (trpE, trpD, trpC, trpB, and trpA) that encode enzymes for tryptophan biosynthesis. These genes are transcribed as a single...
2.2K
Inducible Operons: lac Operon01:25

Inducible Operons: lac Operon

2.5K
The lac operon in Escherichia coli is a model for understanding inducible gene regulation and metabolic flexibility. It integrates local control by lactose and global regulation through catabolite repression, enabling E. coli to preferentially metabolize glucose when available and switch to lactose utilization when glucose is scarce.Structure and Function of the lac OperonThe lac operon contains three structural genes: lacZ (β-galactosidase), lacY (lactose permease), and lacA...
2.5K
Operon Model01:23

Operon Model

1.9K
The operon model represents a fundamental mechanism of gene regulation in prokaryotes, enabling coordinated expression of genes involved in related metabolic or functional pathways. Operons consist of structural genes, a promoter, and an operator, with transcription regulated by repressors, activators, and small effector molecules.Structure and Function of OperonsAn operon is a cluster of structural genes transcribed together under the control of a single promoter. The promoter region...
1.9K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

7.3K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
7.3K
Prokaryotic Transcriptional Activators and Repressors01:58

Prokaryotic Transcriptional Activators and Repressors

26.0K
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.
Transcription of prokaryotic...
26.0K

You might also read

Related Articles

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

Sort by
Same author

Pharmaceuticals, personal care-products and current-use pesticides: a review of the available data from European seas.

Environmental research·2026
Same author

Access to and engagement with healthcare services among women with Children's Social Care involvement during the perinatal period who subsequently died: a confidential enquiry.

BMJ public health·2026
Same author

A step towards microlitter risk assessment: modelling microlitter storage potential of the UK seabed.

Philosophical transactions. Series A, Mathematical, physical, and engineering sciences·2025
Same author

Characteristics, outcomes, and maternity care experiences of women with children's social care involvement who subsequently died: national cohort study and confidential enquiry.

BMJ medicine·2025
Same author

Sedimentary blue carbon around decommissioned oil and gas platforms in the North Sea.

Marine pollution bulletin·2025
Same author

Chemical emissions from offshore wind farms: From identification to challenges in impact assessment and regulation.

Marine pollution bulletin·2025
Same journal

A Video Protocol of a Randomized Controlled Clinical Trial - Electrochemotherapy of Cutaneous Metastases with Reduced Dose Bleomycin (BLESS Trial).

Journal of visualized experiments : JoVE·2026
Same journal

A Standardized Ex Vivo Porcine Oromucosal Model for Evaluating Peptide Fluxes.

Journal of visualized experiments : JoVE·2026
Same journal

Lightweight English Text Classification with Deep Learning Based on Complex System Theory.

Journal of visualized experiments : JoVE·2026
Same journal

Integrating Artificial Intelligence-Assisted Translation Support into English Courses: Effects on Translation Accuracy, Perceived Stress, and Anxiety.

Journal of visualized experiments : JoVE·2026
Same journal

A Toxin-Based Counter-Selection System for Markerless Gene Deletion and High-Density Tn5 Transposon Mutagenesis in Pectobacterium brasiliense.

Journal of visualized experiments : JoVE·2026
Same journal

Seamless Multimodal Human-Robot Communication: Integration Techniques in Human-Computer Interaction.

Journal of visualized experiments : JoVE·2026
See all related articles

Related Experiment Video

Updated: Mar 15, 2026

Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System
11:19

Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System

Published on: August 21, 2016

9.6K

Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System.

Karla A Mettrick1, Nikki Lawrence1, Claire Mason1

  • 1School of Environmental and Life Sciences, University of Newcastle.

Journal of Visualized Experiments : Jove
|September 2, 2016
PubMed
Summary
This summary is machine-generated.

This study introduces a novel method using a Fluorescence Repressor Operator System (FROS) to site-specifically stall DNA replication forks in E. coli. This system allows for detailed visualization and analysis of replication fork collapse and repair mechanisms in living cells.

More Related Videos

A Fluorescence-based Method to Study Bacterial Gene Regulation in Infected Tissues
07:10

A Fluorescence-based Method to Study Bacterial Gene Regulation in Infected Tissues

Published on: February 19, 2019

9.6K
Engineering Adherent Bacteria by Creating a Single Synthetic Curli Operon
15:28

Engineering Adherent Bacteria by Creating a Single Synthetic Curli Operon

Published on: November 16, 2012

15.1K

Related Experiment Videos

Last Updated: Mar 15, 2026

Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System
11:19

Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System

Published on: August 21, 2016

9.6K
A Fluorescence-based Method to Study Bacterial Gene Regulation in Infected Tissues
07:10

A Fluorescence-based Method to Study Bacterial Gene Regulation in Infected Tissues

Published on: February 19, 2019

9.6K
Engineering Adherent Bacteria by Creating a Single Synthetic Curli Operon
15:28

Engineering Adherent Bacteria by Creating a Single Synthetic Curli Operon

Published on: November 16, 2012

15.1K

Area of Science:

  • Molecular Biology
  • Genetics
  • Microbiology

Background:

  • DNA replication machinery can stall due to obstacles like proteins and lesions, leading to replication fork collapse.
  • Replication fork collapse necessitates cellular mechanisms for recovery to ensure accurate DNA duplication and cell division.
  • Previous studies on replication repair pathways often used UV damage, lacking site-specificity.

Purpose of the Study:

  • To develop a site-specific method for inducing and studying replication fork stalling and collapse in Escherichia coli.
  • To enable visualization of replication status in single living cells and analysis of replication intermediates.
  • To investigate the roles of recombination proteins and helicases in replication fork recovery.

Main Methods:

  • Utilized a Fluorescence Repressor Operator System (FROS) to create site-specific protein blocks on DNA.
  • Employed fluorescence microscopy for real-time visualization of replication status in single living cells.
  • Applied 2-dimensional agarose gel electrophoresis for analyzing DNA replication intermediates.
  • Incorporated temperature-sensitive mutants (e.g., DnaBts) for synchronous replication fork collapse induction.
  • Generated genetic knockouts of recombination proteins and helicases to study their roles.

Main Results:

  • Successfully established a system for site-specific induction of replication fork stalling and collapse.
  • Demonstrated visualization of replication dynamics and analysis of replication intermediates in living E. coli.
  • Enabled synchronous collapse of replication forks using temperature-sensitive mutants.
  • Provided a platform for genetic analysis of proteins involved in replication fork repair.

Conclusions:

  • The FROS-based system offers a powerful tool for studying DNA replication fork dynamics and repair in a site-specific manner.
  • This methodology facilitates the investigation of essential cellular mechanisms for maintaining genome stability.
  • The system is adaptable for studying various factors influencing replication fork integrity and recovery.