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Optimized vectors for genetic engineering of Aureobasidium pullulans.

Analeigha V Colarusso¹, Audrey M Williams², Amy S Gladfelter²

¹Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139.

Molecular Biology of the Cell

|April 9, 2025

Summary

This summary is machine-generated.

Aureobasidium pullulans, a versatile black yeast, can now be studied more easily. Researchers developed new genetic tools, including novel vectors, to advance research on cell adaptation and shape regulation in this promising model organism.

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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.