Optimized vectors for genetic engineering of Aureobasidium pullulans

Area of Science:

• Microbiology
• Molecular Biology
• Biotechnology

Background:

• Aureobasidium pullulans is a polyextremotolerant black yeast with significant morphological plasticity.
• Its adaptability makes it a promising model for studying cell adaptation and shape regulation.
• Existing genetic tools for A. pullulans require enhancement to facilitate broader research applications.

Purpose of the Study:

• To expand the genetic toolkit for Aureobasidium pullulans.
• To design and validate a series of novel expression vectors for this yeast.
• To enable advanced investigations into cellular mechanisms and gene function.

Main Methods:

• Design and construction of 25 new vectors.
• Incorporation of seven codon-optimized fluorophores and three selection cassettes.
• Testing of vectors for dual protein expression (GFP and mCherry) and homology-based gene manipulation (deletion and tagging) at the URA3 locus.

Main Results:

• Successfully developed and tested a versatile series of 25 vectors for A. pullulans.
• Demonstrated dual-color fluorescence protein expression and efficient homology-based gene editing.
• Created vectors allowing for endogenous gene tagging and deletion without requiring molecular cloning.

Conclusions:

• The developed vector series significantly enhances the research capabilities for Aureobasidium pullulans.
• These tools will accelerate studies on cell adaptation, morphological plasticity, and gene function in this emerging model system.
• Facilitates a wider range of molecular and cellular experiments in A. pullulans research.