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Transposon tagging using Activator (Ac) in maize.

Thomas P Brutnell1, Liza J Conrad

  • 1Boyce Thompson Institute, Cornell University, Ithaca, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 23, 2003
PubMed
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Activator (Ac) transposon use in maize is limited by low transposition rates. Distributing Ac elements throughout the genome and utilizing their linked site preference offers a strategy to overcome this for gene isolation.

Area of Science:

  • Plant genetics
  • Molecular biology
  • Genomics

Background:

  • The Activator (Ac) transposable element is a valuable tool for gene isolation in plants.
  • Its application in maize is hindered by a low germinal transposition rate.

Purpose of the Study:

  • To detail a system for utilizing Ac elements in maize for gene isolation.
  • To provide a protocol for cloning DNA flanking Ac insertions.

Main Methods:

  • Genetic manipulation of Ac element distribution in the maize genome.
  • Leveraging Ac's tendency to transpose to linked sites.
  • Development of a protocol for flanking sequence isolation.

Main Results:

  • A strategy to enhance Ac transposition efficiency in maize was developed.

Related Experiment Videos

  • The method allows for targeted gene isolation and characterization.
  • A detailed protocol for cloning flanking DNA is presented.

    • Conclusions:

    • Genomic distribution of Ac elements can overcome low transposition rates in maize.
    • This approach facilitates gene isolation and characterization in its native host.
    • The provided protocol enables efficient cloning of DNA adjacent to Ac insertions.