T-DNA insertion, plasmid rescue and integration analysis in the model mycorrhizal fungus Laccaria bicolor.
Bottom Line: Our results demonstrate that the plasmid rescue approach can be used for resolving T-DNA integration sites in Laccaria.Neither obvious sequence similarities were found between these sites and the T-DNA borders indicating non-homologous integration of the transgenes.Agrobacterium-mediated gene transfer is a powerful tool that can be used for functional gene studies in Laccaria and will be helpful along with plasmid rescue in searching for relevant fungal genes involved in the symbiotic process.
Affiliation: Laboratorio de Micología Molecular, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, (B1876BXD) Bernal, Provincia de Buenos Aires, Argentina.Show MeSH
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Mentions: None of the analysed dikaryotic fungal transgenic strains with a T‐DNA insertion showed an obvious altered phenotype such as hyphal morphology or growth rate when cultivated on P5 media. Three transgenic strains (4, 24 and 36, Table 1) with an interruption in a predicted gene were selected in order to test by reverse transcription polymerase chain reaction (RT‐PCR) whether a single integration in one of the gene copies of the dikaryotic strain can cause a detectable reduction on the gene expression level or not (Fig. 5).
Affiliation: Laboratorio de Micología Molecular, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, (B1876BXD) Bernal, Provincia de Buenos Aires, Argentina.