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

Mutations in the mariner transposase: the D,D(35)E consensus sequence is nonfunctional

A R Lohe1, D De Aguiar, D L Hartl

  • 1Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, MA 02138, USA.

Proceedings of the National Academy of Sciences of the United States of America
|February 18, 1997
PubMed
Summary

A new Drosophila melanogaster eye-color screen simplifies studying mariner transposase mutations. This research identified critical residues for DNA excision and revealed dominant-negative effects, advancing transposon research.

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

AVERAGE FITNESS OF POPULATIONS OF DROSOPHILA MELANOGASTER AS ESTIMATED USING COMPOUND-AUTOSOME STRAINS.

Evolution; international journal of organic evolution·2017
Same author

ESTIMATION OF AVERAGE FITNESS OF POPULATIONS OF DROSOPHILA MELANOGASTER AND THE EVOLUTION OF FITNESS IN EXPERIMENTAL POPULATIONS.

Evolution; international journal of organic evolution·2017
Same author

ANALYSIS OF A GENERAL POPULATION GENETIC MODEL OF MEIOTIC DRIVE.

Evolution; international journal of organic evolution·2017
Same author

Population dynamics of sperm and pollen killers.

TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·2014
Same author

Natural variation of the Y chromosome suppresses sex ratio distortion and modulates testis-specific gene expression in Drosophila simulans.

Heredity·2013
Same author

Estimation of inbreeding by random walks in pedigrees.

TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·2011

Area of Science:

  • Molecular Biology
  • Genetics
  • Drosophila melanogaster research

Background:

  • Studying eukaryote transposases is challenging due to difficulties in isolating mutations.
  • Mariner transposases are mobile genetic elements found across diverse organisms.
  • Understanding transposase function is crucial for gene regulation and genome stability.

Purpose of the Study:

  • To develop a facile method for isolating and analyzing mutations in the Drosophila melanogaster mariner transposase.
  • To identify key amino acid residues essential for the in vivo excision activity of the mariner transposase.
  • To investigate the functional interactions between wild-type and mutant transposase proteins.

Main Methods:

  • Developed a novel eye-color screening system in Drosophila melanogaster for identifying transposase mutants.

Related Experiment Videos

  • Utilized ethyl methanesulfonate mutagenesis and site-directed mutagenesis to generate transposase variants.
  • Assayed transposase activity by measuring germline excision frequency of a target mariner element in heterozygous genotypes.

    • Main Results:

    • Identified 18 critical residues essential for in vivo excision of a target mariner element.
    • Over half of the mutant transposase proteins exhibited dominant-negative effects, inhibiting wild-type activity.
    • Replacing the D,D(34)D domain with a D,D(34)E consensus sequence abolished transposase function.

    Conclusions:

    • The developed eye-color screen is effective for studying mariner transposase mutations in Drosophila.
    • Specific residues within the D,D(34)D motif are crucial for transposase activity and suggest subunit interactions.
    • Mutations in the initiation codon and nuclear localization signal also eliminate transposase function.