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 Experiment Videos

Restriction enzyme-mediated integration (REMI) mutagenesis.

Adam Kuspa1

  • 1Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 8, 2006
PubMed
Summary

Restriction enzyme-mediated integration (REMI) efficiently inserts linear DNA into the Dictyostelium genome. This method facilitates genetic screens and gene discovery by enabling targeted DNA integration with minimal genomic damage.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Loss of the Polyketide Synthase StlB Results in Stalk Cell Overproduction in Polysphondylium violaceum.

Genome biology and evolution·2020
Same author

(Auto)Biographical reflections on the contributions of William F. Loomis (1940-2016) to Dictyostelium biology.

The International journal of developmental biology·2019
Same author

Microbiome management in the social amoeba Dictyostelium discoideum compared to humans.

The International journal of developmental biology·2019
Same author

Social amoebae establish a protective interface with their bacterial associates by lectin agglutination.

Science advances·2019
Same author

Cooperative predation in the social amoebae Dictyostelium discoideum.

PloS one·2019
Same author

Lectins modulate the microbiota of social amoebae.

Science (New York, N.Y.)·2018

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Efficient and targeted DNA integration is crucial for genetic manipulation in model organisms.
  • The Dictyostelium discoideum genome presents unique challenges for stable DNA insertion.
  • Previous methods lacked precision and could cause significant genomic disruption.

Purpose of the Study:

  • To describe a novel method for integrating linear DNA into the Dictyostelium genome.
  • To establish a reliable technique for genetic screens and marker placement.
  • To enable efficient gene discovery through targeted mutagenesis.

Main Methods:

  • Restriction enzyme-mediated integration (REMI) using linearized plasmid DNA and a specific restriction enzyme.
  • Transformation of Dictyostelium cells with the DNA and enzyme mixture.

Related Experiment Videos

  • Stimulation of DNA integration into compatible restriction sites within the host genome.

    • Main Results:

    • REMI facilitates the integration of linear DNA into the Dictyostelium genome.
    • Integration typically occurs as a single-copy event with minimal collateral genomic damage.
    • The method has been successfully used to identify numerous genes based on mutant phenotypes.

    Conclusions:

    • REMI is a powerful and efficient tool for genetic manipulation in Dictyostelium.
    • This technique significantly aids in genetic screening and the precise localization of genetic markers.
    • REMI has proven invaluable for gene discovery and functional genomics studies in Dictyostelium.