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Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

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...

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Related Experiment Video

Updated: Sep 6, 2025

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

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Published on: January 8, 2015

Lambda Red Recombineering in Shigella flexneri.

Adrian Herod¹, Julie Ryu¹, John Rohde²

¹Department of Microbiology and Immunology, Dalhousie University Halifax, Halifax, NS, Canada.

Methods in Molecular Biology (Clifton, N.J.)

|June 27, 2022

Summary

This summary is machine-generated.

Developing vaccines for shigellosis, a deadly diarrheal disease, is challenging. This study presents a new method for creating gene deletions in Shigella flexneri, aiding in understanding disease mechanisms and vaccine development.

Keywords:

Congo red Conjugation Shigella Vaccine development λ-Red recombination

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Last Updated: Sep 6, 2025

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Author Spotlight: Advancements in Understanding and Combatting Shigella Infections

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Published on: February 9, 2024

Area of Science:

Microbiology
Infectious Diseases
Vaccinology

Background:

Shigellosis is a significant global health problem causing severe diarrhea and mortality.
Developing effective vaccines against Shigella is hindered by poor understanding of disease mechanisms and genetic manipulation challenges.
Precise gene deletion protocols exist but are not widely adopted for Shigella flexneri mutant construction.

Purpose of the Study:

To describe a streamlined method for constructing gene deletions in Shigella flexneri.
To facilitate the study of Shigella pathogenesis and the development of novel vaccines.
To provide researchers with practical tools for genetic manipulation of Shigella.

Main Methods:

Utilizing the λ-red recombination system for precise gene deletions.
Employing laboratory-developed tools to enhance the efficiency of genetic manipulation.
Outlining specific steps for constructing deletion mutants in Shigella flexneri.

Main Results:

Successfully established a reliable protocol for creating gene deletions in Shigella flexneri.
Developed and validated laboratory tools that simplify the construction of mutants.
Demonstrated the utility of the λ-red recombination system for Shigella research.

Conclusions:

The described λ-red recombination method offers a practical approach for generating Shigella flexneri deletion mutants.
This methodology will aid in elucidating Shigella pathogenesis and advancing vaccine research.
The developed tools provide a valuable resource for the scientific community studying shigellosis.