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Homologous Recombination

Homologous Recombination

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...

Homologous Recombination

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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...

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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.
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Related Experiment Video

Updated: May 15, 2026

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Published on: September 11, 2022

IntDOT interactions with core sites during integrative recombination.

Jennifer Laprise¹, Sumiko Yoneji, Jeffrey F Gardner

¹Department of Microbiology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA. jlapri2@illinois.edu

Journal of Bacteriology

|January 22, 2013

Summary

Integrative and conjugative elements (ICEs) spread antibiotic resistance. Researchers studied CTnDOT

Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

Integrative and conjugative elements (ICEs) contribute to the spread of antibiotic resistance genes.
CTnDOT is a well-studied ICE in Bacteroides spp., carrying resistance genes and a tyrosine recombinase (IntDOT) for chromosomal integration/excision.
ICE integration occurs at specific attB sites, involving core sites like B, B', D, and D'.

Purpose of the Study:

To investigate the role of conserved sequences in attB sites for CTnDOT integration.
To characterize the interaction between the IntDOT recombinase and specific base pairs within the attB site.

Main Methods:

Mutational analysis of attB sites to identify critical sequences for integration.
Base analog studies to probe IntDOT's interaction with DNA bases.

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

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

Published on: June 25, 2013

Related Experiment Videos

Last Updated: May 15, 2026

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Published on: September 11, 2022

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

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

Published on: June 25, 2013

Main Results:

Identified crucial T-A base pairs at positions -5 (B and D sites) and +5 (B' site) for integrative recombination.
Base analog studies suggest IntDOT specifically contacts adenine (A) residues in the major groove at positions -5 and +5.
IntDOT's interaction with the adenine at position -5 in the B core site is essential for initiating the first DNA strand exchange.

Conclusions:

The conserved B core site sequence is vital for CTnDOT integration.
Specific base pair interactions, particularly with adenine, are critical for IntDOT-mediated recombination.
Understanding these interactions provides insight into the mechanism of ICE integration.