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Published on: February 14, 2020
A hyperactive transposase of the maize transposable element activator (Ac)
Katina Lazarow1, My-Linh Du, Ruth Weimer
1Dahlem Centre of Plant Sciences, Freie Universität Berlin, 14195 Berlin, Germany.
A hyperactive transposase derivative, AcTPase(4x), significantly increases Ds element excisions in yeast and Arabidopsis. This discovery enhances Ac/Ds-based tools for plant insertional mutagenesis and gene isolation.
Area of Science:
- Genetics
- Molecular Biology
- Plant Science
Background:
- Activator/Dissociation (Ac/Ds) transposable elements are crucial for insertional mutagenesis in plants.
- Low transposition frequencies limit Ac/Ds utility in some plant species.
Purpose of the Study:
- To develop a hyperactive Ac transposase derivative for enhanced Ac/Ds-based mutagenesis.
- To evaluate the excision and insertion characteristics of the novel AcTPase(4x).
Main Methods:
- Engineering a hyperactive Ac transposase derivative (AcTPase(4x)).
- Assessing Ds excision and reintegration frequencies in yeast (Saccharomyces cerevisiae).
- Evaluating AcTPase(4x) activity in Arabidopsis thaliana.
Main Results:
- AcTPase(4x) demonstrated a 100-fold increase in Ds excisions in yeast compared to wild-type.
- Ds reintegration frequency remained unchanged (57%) with AcTPase(4x).
- AcTPase(4x) showed a >6-fold increase in Ds excision in Arabidopsis, with insertion biases similar to wild-type but lacking specific site preferences.
Conclusions:
- AcTPase(4x) significantly enhances Ac/Ds transposition efficiency.
- This hyperactive transposase derivative offers improved utility for plant insertional mutagenesis strategies.
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