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United States Department of Agriculture

Agricultural Research Service

Title: Development of a Remi-Transformation Procedure for the Mutational Tagging of Genes in Paecilomyces Fumosoroseus

item Cantone, Frank
item Vandenberg, John

Submitted to: Fungal Genetics Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: March 15, 1997
Publication Date: N/A

Technical Abstract: We have developed a transformation system for the entomopathogenic fungus, Paecilomyces fumosoroseus, that relies on restriction enzyme-mediated integration, under the selection of bialaphos. This procedure is being used as a tool for the mutational tagging of genes and the creation of transformants deficient in some aspect of pathogenicity. Depending upon the enzyme and vector used transformation frequencies were either the enzyme and vector used transformation frequencies were either unaffected or increased versus non-REMI treatments. Restriction mapping and Southern hybridization analyses of transformant DNAs revealed that plasmids inserted randomly throughout the genome. However, results also revealed that integrations occurred other than by a simple ligation-repair model. From the pattern of REMI events we were unable to regenerate the original restriction sites of the plasmid with the enzyme initially used in the transformation procedure. Instead we observed that one or both sites were destroyed during the integration of the plasmid in the genome. We have currently obtained several pigmentation and pathogenicity mutants using this transformation procedure.

Last Modified: 4/22/2015