Drug-resistant cassettes for the efficient transformation of Candida guilliermondii wild-type strains

• DOI: 10.1111/j.1567-1364.2011.00731.x
• Publication Date: 2011-04-13T20:08:13Z
• Authors: Yoann Millerioux; Marc Clastre; Andrew J. Simkin; Vincent Courdavault; Emeline Marais; Andriy A. Sibirny; Thierry Noël; Joël Crèche; Nathalie Giglioli...; Nicolas Papon
• Keywords: Genetics, Microbial; 0301 basic medicine; Antifungal Agents; 3. Good health; Mutagenesis, Insertional; 03 medical and health sciences; [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology; Transformation, Genetic; Drug Resistance, Fungal; Streptothricins; Hygromycin B; Selection, Genetic; [SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology; Candida

Candida guilliermondii is an opportunistic emerging fungal agent of candidiasis often associated with oncology patients. This yeast also remains an interesting biotechnological model for the industrial production of value-added metabolites. The recent whole-genome sequencing of the C. guilliermondii ATCC 6260 reference strain provides an interesting resource for elucidating new molecular events supporting pathogenicity, antifungal resistance and for exploring the potential of yeast metabolic engineering. In the present study, we designed an efficient transformation system for C. guilliermondii wild-type strains using both nourseothricin- and hygromycin B-resistant markers. To demonstrate the potential of these drug-resistant cassettes, we carried out the disruption and the complementation of the C. guilliermondii FCY1 gene (which encodes cytosine deaminase) known to be associated with flucytosine sensitivity in yeast. These two new dominant selectable markers represent powerful tools to study the function of a large pallet of genes in this yeast of clinical and biotechnological interest.