Methylation of the foreign transposon Restless in vegetative mycelia of Neurospora crassa

Frank Windhofer, David E.A. Catcheside, Frank Kempken

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    16 Citations (Scopus)

    Abstract

    Methylation of foreign and/or repeated sequences in the filamentous fungus Neurospora crassa is believed to be directed against invading transposable elements. To test this hypothesis, the fate of a transposon in N. crassa was investigated. Vectors were constructed which carried the transposon Restless, an active class-II element isolated from the fungus Tolypocladium inflatum. These vectors were introduced into N. crassa strains by protoplast transformation. Two strategies were employed: (1) ectopic multi-copy integration, and (2) site-specific single-copy integration at the his-3 locus. All ectopic transformants exhibited strong methylation as confirmed by Southern hybridization of genomic DNA digested with the methylation-sensitive endonuclease Sau3AI and the methylation-insensitive endonuclease NdeII. Single copies of Restless integrated at the his-3 locus were not methylated. These results are discussed with respect to non-RIP methylation and potential consequences for gene-tagging strategies based on the use of Restless.

    Original languageEnglish
    Pages (from-to)194-199
    Number of pages6
    JournalCurrent Genetics
    Volume37
    Issue number3
    DOIs
    Publication statusPublished - Mar 2000

    Keywords

    • DNA methylation
    • Neurospora crassa
    • Transposon Restless

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