Optimized vectors for genetic engineering of Aureobasidium pullulans

ABSTRACT Aureobasidium pullulans is a polyextremotolerant black yeast that exhibits impressive morphological plasticity. Consequently, it shows promise as a model system for investigating mechanisms of cell adaptation to different environments and the regulation of cell shape. Here, we build upon the current toolkit for working with A. pullulans and design and test 25 vectors with seven different codon-optimized fluorophores and three selection cassettes. This includes vectors that allow for dual expression of GFP and mCherry tagged proteins at the URA3 locus and vectors that enable homology-based deletion or C-terminal tagging of endogenous genes without the need for cloning. This versatile vector series for working with A. pullulans will enable a broad range of experiments in this emerging model system. SUMMARY An optimized plasmid toolkit for genetic engineering of the emerging model fungus, A. pullulans. Competing Interest Statement The authors have declared no competing interest. ABBREVIATIONS - DNA - Deoxyribonucleic acid - doLEU - media lacking leucine - doURA - media lacking uracil - G418 - Geneticin - Hyg - hygromycin B - MIC - minimal inhibitory concentration - mNG - mNeonGreen - Nat - nourseothricin