Overcoming recalcitrant transformation and gene manipulation in Pucciniomycotina yeasts
Erika P Abbott, Giuseppe Ianiri, Raffaello Castoria, Alexander Idnurm
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY | SPRINGER | Published : 2013
The red yeasts of the Pucciniomycotina have rarely been transformed with DNA molecules. Transformation methods were recently developed for a species of Sporobolomyces, based on selection using uracil auxotrophs and plasmids carrying the wild-type copies of the URA3 and URA5 genes. However, these plasmids were ineffective in the transformation of closely related species. Using the genome-sequenced strain of Rhodotorula graminis as a starting point, the URA3 and URA5 genes were cloned and tested for the transformation ability into different Pucciniomycotina species by biolistic and Agrobacterium-mediated transformations. Transformation success depended on the red yeast species and the origin o..View full abstract
Awarded by United States National Science Foundation
Awarded by Italian Ministry of Education, University and Scientific Research
Awarded by Italian Ministry of Foreign Affairs
We thank Joseph Heitman, James Fraser, and the FGSC for providing strains. This research was supported by grants from the United States National Science Foundation (MCB-0920581), the Italian Ministry of Education, University and Scientific Research (PRIN 2008, 2008JKH2MM), and the Italian Ministry of Foreign Affairs (joint research project LS-7, within the executive programme of cooperation in the field of science and technology between Italy and USA 2008-2010). G. I. was also supported by a scholarship from the Department of Agricultural, Environmental and Food Sciences, University of Molise, Italy.