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Transforming the sapstaining fungus Ophiostoma piceae with Agrobacterium tumefaciens

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Abstract:

A transformation protocol mediated by Agrobacterium tumefaciens is described for the sapstaining fungus Ophiostoma piceae. We compared transformants obtained from Agrobacterium with those obtained from yeast-like cells made into spheroplasts and treated with CaCl2. For all putative transformants analyzed, Southern hybridization confirmed that the hygromycin resistance gene had been integrated into the genomic DNA. While all transformants obtained from the treated spheroplasts had multiple copy vector insertion, 85% of the Agrobacterium-mediated transformants had single copy vector insertion.Key words: Ophiostoma piceae, genetic transformation, Ti plasmid.

Nous faisons la description d'un protocole de transformation facilitée par Agrobacterium tumefaciens pour le champignon Ophiostoma piceae responsable de la coloration de l'aubier. Nous avons comparé les transformants obtenus avec Agrobacterium avec ceux de cellules lévuriformes changées en sphéroplastes et traitées avec du chlorure de calcium. Chez tous les transformants présumés, une hybridation de type Southern a confirmé que le gène de résistance à l'hygromycine s'était intégré dans l'ADN génomique. Alors que tous les transformants obtenus à partir des sphéroplastes renfermaient des copies multiples du vecteur inséré, 85 % des transformants facilités par Agrobacterium avaient une copie unique du vecteur inséré.Mots clés : Ophiostoma piceae, transformation génétique, plasmide Ti.[Traduit par la Rédaction]

Document Type: Research Article

Publication date: March 1, 2003

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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