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Related Experiment Videos

Analysis of extrachromosomal Ac/Ds transposable elements.

V Gorbunova1, A A Levy

  • 1Plant Sciences Department, The Weizmann Institute of Science, Rehovot, 76100 Israel.

Genetics
|May 3, 2000
PubMed
Summary

Investigating maize Ac/Ds element transposition, this study reveals extrachromosomal circular and linear products. Analysis suggests the first flanking nucleotide at excision sites resembles the donor sequence, offering insights into transposition mechanisms.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Plant Science

Background:

  • The transposition mechanism of maize Activator/Dissociation (Ac/Ds) elements remains unclear.
  • Distinguishing between excision models involving different staggered cuts and hairpin formation has been challenging.
  • True transposition intermediates have not been identified.

Purpose of the Study:

  • To investigate the excision mechanism of maize Ac/Ds elements.
  • To analyze extrachromosomal excision products for insights into transposition intermediates.
  • To differentiate between proposed models of Ac/Ds element excision.

Main Methods:

  • Plasmid rescue was employed to recover and analyze extrachromosomal excision products.
  • Sequencing of circular and linear extrachromosomal Ds elements was performed.

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  • Nucleotide sequences at the termini of excision products were compared to the original donor site sequences.

    • Main Results:

    • Ds excision was shown to be associated with the formation of extrachromosomal circular molecules.
    • Evidence for the formation of linear extrachromosomal species during Ds excision was presented.
    • Analysis of nucleotide sequences at excision termini indicated nucleotide additions/deletions, suggesting these are not true intermediates.
    • The first nucleotide adjacent to the transposon end showed significant similarity to the donor site's flanking nucleotide, while subsequent nucleotides did not.

    Conclusions:

    • Extrachromosomal circular and linear products are formed during Ds excision.
    • The observed sequence similarities at excision termini provide crucial data for refining models of Ac/Ds transposition.
    • Findings suggest a specific mechanism for the initial step of Ds element excision.