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

116
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...
116
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.1K
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...
6.1K
Viral Recombination00:57

Viral Recombination

23.7K
Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.
23.7K
Homologous Recombination02:31

Homologous Recombination

50.8K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
50.8K
Recombinant DNA01:09

Recombinant DNA

96.2K
Overview
96.2K
Replication in Prokaryotes01:32

Replication in Prokaryotes

25.1K
DNA replication has three main steps: initiation, elongation, and termination. Replication in prokaryotes begins when initiator proteins bind to the single origin of replication (ori) on the cell's circular chromosome. Replication then proceeds around the entire circle of the chromosome in each direction from the two replication forks, resulting in two DNA molecules.
Many Proteins Work Together to Replicate the Chromosome
Replication is coordinated and carried out by a host of specialized...
25.1K

You might also read

Related Articles

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

Sort by
Same author

Enhancement of RecET-mediated in vivo linear DNA assembly by a xonA mutation.

PloS one·2026
Same author

A λ Phage Platform for Successful Therapeutic Display of Protein Antigens.

bioRxiv : the preprint server for biology·2026
Same author

Correction: Recombineering: Genetic Engineering in Escherichia coli Using Homologous Recombination.

Current protocols·2024
Same author

Stability and gene strand bias of lambda prophages and chromosome organization in <i>Escherichia coli</i>.

mBio·2024
Same author

The crystal structure of bacteriophage λ RexA provides novel insights into the DNA binding properties of Rex-like phage exclusion proteins.

Nucleic acids research·2024
Same author

Everything OLD is new again: How structural, functional, and bioinformatic advances have redefined a neglected nuclease family.

Molecular microbiology·2023
Same journal

A Novel Laboratorial Approach to Evaluate Bacterial Microleakage of Endodontic Sealers.

Current protocols·2026
Same journal

TRIAGE Toolkit: Streamlined Discovery of Regulatory Genes and Elements.

Current protocols·2026
Same journal

High-throughput Profiling of Pseudouridines in Microbiome-derived Bacterial RNA.

Current protocols·2026
Same journal

Recombinant Protein Expression in Rhodococcus species.

Current protocols·2026
Same journal

Streamlined In Vitro Transcription for Generating Self-Amplifying RNA With Modified Nucleotides.

Current protocols·2026
Same journal

CODEC Library Preparation From Genomic DNA.

Current protocols·2026
See all related articles

Related Experiment Video

Updated: Aug 16, 2025

Recombineering Homologous Recombination Constructs in Drosophila
14:23

Recombineering Homologous Recombination Constructs in Drosophila

Published on: July 13, 2013

19.4K

Recombineering in Non-Model Bacteria.

Anna Corts1, Lynn C Thomason2, Nina Costantino2

  • 1Cultivarium, Watertown, Massachusetts, USA.

Current Protocols
|December 22, 2022
PubMed
Summary
This summary is machine-generated.

Recombineering enables precise in vivo genetic engineering in diverse bacteria using homologous recombination. This study details applications, challenges, and new systems for single-strand DNA (ssDNA) and double-strand DNA (dsDNA) recombineering in non-model organisms.

Keywords:
RecETannealasenon-model bacteriarecombineeringλ Red

More Related Videos

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
09:02

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

16.6K
Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy
11:40

Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy

Published on: June 25, 2013

12.2K

Related Experiment Videos

Last Updated: Aug 16, 2025

Recombineering Homologous Recombination Constructs in Drosophila
14:23

Recombineering Homologous Recombination Constructs in Drosophila

Published on: July 13, 2013

19.4K
Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
09:02

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

16.6K
Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy
11:40

Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy

Published on: June 25, 2013

12.2K

Area of Science:

  • Microbiology
  • Molecular Biology
  • Synthetic Biology

Background:

  • Recombineering, a method for in vivo genetic engineering, utilizes bacteriophage-encoded homologous recombination proteins.
  • It allows precise genomic DNA modification at short homology sites (35-50 nt) without restriction enzyme dependence.
  • The technology employs linear DNA substrates like PCR products, dsDNA, or ssDNA, introduced via electroporation.

Purpose of the Study:

  • To review applications, challenges, and factors influencing ssDNA and dsDNA recombineering in various non-model bacteria.
  • To highlight recent breakthroughs and new homologous recombineering systems from non-model bacteria.
  • To provide guidance for researchers on selecting or developing recombineering systems for specific microbes.

Main Methods:

  • Description of established phage λ Red and Rac RecET systems used in diverse bacteria.
  • Introduction of novel homologous ssDNA and dsDNA recombineering systems from non-model bacteria.
  • Detailed protocols for ssDNA recombineering in Shewanella species, including CRISPR/Cas9 as a counter-selection tool.

Main Results:

  • Successful application of recombineering in a wide range of Gram-negative and Gram-positive non-model bacteria.
  • Development and characterization of new ssDNA and dsDNA recombineering systems.
  • Demonstration of CRISPR/Cas9 as an effective counter-selection method to enhance recombinant recovery.

Conclusions:

  • Recombineering is a versatile tool for precise genome engineering in diverse bacterial species.
  • New systems and strategies, including oligo-mediated recombination, expand the utility of recombineering in undomesticated bacteria.
  • This work provides a decision-making framework to aid researchers in applying or developing recombineering strategies for their specific microbial targets.