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Related Experiment Videos

Tagged Transcriptome Display (TTD) in indica rice using Ac transposition.

A Kohli1, J Xiong, R Greco

  • 1Molecular Biotechnology Unit, John Innes Centre, Norwich Research Park, UK.

Molecular Genetics and Genomics : MGG
|October 9, 2001
PubMed
Summary

Researchers created transgenic rice with the Activator transposon (Ac) to study its movement. They developed a Tagged Transcriptome Display (TTD) to identify genes activated by transposon insertions, offering a new tool for gene discovery in plants.

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Area of Science:

  • Plant molecular biology
  • Genetics and genomics
  • Transposable element research

Background:

  • Transposable elements (TEs) are mobile DNA sequences that can alter gene function and genome structure.
  • Understanding TE behavior in heterologous systems is crucial for gene discovery and crop improvement.
  • The Activator (Ac) transposon from maize is a well-characterized system for genetic studies.

Purpose of the Study:

  • To develop and characterize transgenic indica rice lines harboring the maize Activator (Ac) transposon.
  • To investigate the transposition dynamics and insertion patterns of Ac in rice.
  • To establish a novel method for identifying genes tagged by Ac insertions in transcribed sequences.

Main Methods:

  • Generation of transgenic indica rice lines with the Ac transposon and a GFP reporter.

Related Experiment Videos

  • Monitoring Ac excision events using GFP activity in various rice tissues.
  • Molecular analysis of Ac excision and re-insertion events.
  • Development of Tagged Transcriptome Display (TTD) using methylation-sensitive enzymes and cDNA probes.
  • Sequencing of transposon-tagged sites to identify homologous genes/ESTs.

    • Main Results:

    • Ac excision was observed in 60% of transformants, with early and late excision phenotypes.
    • Active transposition and copy number amplification of Ac were detected in progeny lines.
    • The TTD technique successfully identified transposon insertions in transcribed sequences.
    • A significant proportion (12%) of amplified tags showed homology to known genes or ESTs, exceeding random expectations.

    Conclusions:

    • The transgenic rice system effectively harbors and exhibits Ac transposon activity.
    • Tagged Transcriptome Display (TTD) is an efficient approach for functional genomics and gene discovery in rice.
    • This method facilitates the identification of genes involved in specific biological processes through transposon tagging.