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Mobility assays confirm the broad host-range activity of the Minos transposable element and validate new transformation tools

Authors

  • A. G. Klinakis,

    1. Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas (FORTH), Heraklion, Crete, Greece, also at: Department of Biology, University of Crete, Heraklion, Crete, Greece, and
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  • T. G. Loukeris,

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    • *

      Present address: European Molecular Biology Laboratory, EMBL, Meyerhofstr. 1, Heidelberg, Germany.

  • A. Pavlopoulos,

    1. also at: Division of Medical Sciences, Medical School, University of Crete, Heraklion, Crete, Greece
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  • C. Savakis

    Corresponding author
    1. also at: Division of Medical Sciences, Medical School, University of Crete, Heraklion, Crete, Greece
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C. Savakis, IMBB-FORTH, PO Box 1527, Heraklion, Crete, Greece 71110. Tel.: + 30–81–391114; fax: + 30–81–391955; e-mail: savakis@imbb.forth.gr

Abstract

Fast and reliable methods for assessing the mobility of the transposable element Minos have been developed. These methods are based on the detection of excision and insertion of Minos transposons from and into plasmids which are co-introduced into cells. Excision is detected by polymerase chain reaction (PCR) with appropriate primers. Transposition is assayed by marker rescue in Escherichia coli, using a transposon plasmid that carries a tetracycline resistance gene and a target plasmid carrying a gene that can be selected against in E. coli. Using both assays, Minos was shown to transpose in Drosophila melanogaster cells and embryos, and in cultured cells of a mosquito, Aedes aegypti, and a lepidopteran, Spodoptera frugiperda. In all cases, mobility was dependent on the presence of exogenously supplied transposase, and both excision and transposition were precise. The results indicate that Minos can transpose in heterologous insect species with comparable efficiencies and therefore has the potential to be used as a transgenesis vector for diverse species.

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