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Related Experiment Videos

Single-step selection for Ty1 element retrotransposition.

M J Curcio1, D J Garfinkel

  • 1Advanced BioScience Laboratories-Basic Research Program, National Cancer Institute-Frederick Cancer Research and Development Center, MD 21702-1201.

Proceedings of the National Academy of Sciences of the United States of America
|February 1, 1991
PubMed
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Researchers developed a novel reporter system to track yeast retrotransposon Ty1 RNA-mediated transposition. This method allows for the study of retroelements and estimates Ty1 transposition rates, revealing correlations with transcript abundance.

Area of Science:

  • Molecular Biology
  • Genetics
  • Yeast Biology

Background:

  • The yeast retrotransposon Ty1 is a mobile genetic element.
  • Studying retrotransposon activity is crucial for understanding genome dynamics.
  • Existing methods for tracking Ty1 transposition have limitations.

Purpose of the Study:

  • To develop a novel reporter system for studying RNA-mediated transposition of the yeast Ty1 retrotransposon.
  • To enable the investigation of retroelements in other organisms using this reporter system.
  • To quantify the transposition rate of Ty1 elements.

Main Methods:

  • A reporter gene (yeast HIS3 interrupted by an artificial intron, AI) was engineered and inserted into the Ty1 element.
  • The reporter construct (Ty1-H3mHIS3AI) was placed under the control of an inducible GAL1 promoter.

Related Experiment Videos

  • Histidine prototrophy (His+) was used as a selectable marker for successful splicing and retrotransposition events.

    • Main Results:

    • Galactose induction of the Ty1-H3mHIS3AI element led to a high frequency of His+ cells.
    • Spontaneous His+ revertants from genomic Ty1-H3mHIS3AI elements confirmed retrotransposition.
    • The Ty1 transposition rate was estimated to be between 3 x 10(-7) and 1 x 10(-5) per element per generation.
    • Ty1 transposition rates correlated with the relative abundance of their transcripts.

    Conclusions:

    • The developed reporter system is effective for selecting and studying RNA-mediated Ty1 transposition events.
    • This assay provides a quantitative measure of Ty1 transposition rates.
    • Transcript abundance influences Ty1 transposition frequency, offering insights into retroelement regulation.