Methylation of the foreign transposon Restless in vegetative mycelia of Neurospora crassa
| Autor: | Frank Kempken, David E. A. Catcheside, Frank Windhofer |
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| Rok vydání: | 2000 |
| Předmět: |
Genetics
Transposable element Neurospora crassa biology Genetic Vectors fungi DNA Recombinant Crassa General Medicine Methylation DNA Methylation biology.organism_classification Blotting Southern genomic DNA Transformation Genetic Tolypocladium inflatum DNA methylation DNA Transposable Elements DNA Fungal Deoxyribonucleases Type II Site-Specific Southern blot |
| Zdroj: | Current Genetics. 37:194-199 |
| ISSN: | 1432-0983 0172-8083 |
| DOI: | 10.1007/s002940050519 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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