Optimized vectors for genetic engineering of Aureobasidium pullulans

Area of Science:

• Mycology
• Synthetic Biology
• Molecular Biology

Background:

• Aureobasidium pullulans is a polyextremotolerant black yeast known for its morphological plasticity.
• Its adaptability makes it a promising model organism for studying cell adaptation and shape regulation.

Purpose of the Study:

• To expand the genetic toolkit for Aureobasidium pullulans.
• To create versatile vectors for gene expression, tagging, and deletion in A. pullulans.

Main Methods:

• Design and testing of 25 novel vectors.
• Inclusion of seven codon-optimized fluorophores (e.g., GFP, mCherry).
• Development of vectors for dual expression, homology-based gene deletion, and C-terminal tagging without cloning.

Main Results:

• A series of 25 functional vectors for A. pullulans research were successfully created and validated.
• Vectors enable dual protein expression, endogenous gene tagging, and gene deletion.
• The new tools support experiments at the URA3 locus and facilitate homology-based modifications.

Conclusions:

• The developed vector series significantly enhances the experimental capabilities for studying Aureobasidium pullulans.
• This versatile toolkit will accelerate research into cell adaptation and morphogenesis in this emerging model system.