Chemical transformation of the multibudding yeast, Aureobasidium pullulans
Name
jcb_202402114.pdf
Description
Published version
Size
4.46 MB
Format
Adobe PDF
Checksum (MD5)
3dd9b9bdb5c4df0f0f901bfb1a1201b3
Author(s)
Wirshing, Alison CE
Petrucco, Claudia A
Lew, Daniel J
Date Issued
June 27, 2024
Journal
Journal of Cell Biology
Publisher
Rockefeller University Press
Citation
Alison C.E. Wirshing, Claudia A. Petrucco, Daniel J. Lew; Chemical transformation of the multibudding yeast, Aureobasidium pullulans. J Cell Biol 7 October 2024; 223 (10): e202402114.
Version
Final published version
Abstract
Aureobasidium pullulans is a ubiquitous polymorphic black yeast with industrial and agricultural applications. It has recently gained attention amongst cell biologists for its unconventional mode of proliferation in which multinucleate yeast cells make multiple buds within a single cell cycle. Here, we combine a chemical transformation method with genome-targeted homologous recombination to yield ∼60 transformants/μg of DNA in just 3 days. This protocol is simple, inexpensive, and requires no specialized equipment. We also describe vectors with codon-optimized green and red fluorescent proteins for A. pullulans and use these tools to explore novel cell biology. Quantitative imaging of a strain expressing cytosolic and nuclear markers showed that although the nuclear number varies considerably among cells of similar volume, total nuclear volume scales with cell volume over an impressive 70-fold size range. The protocols and tools described here expand the toolkit for A. pullulans biologists and will help researchers address the many other puzzles posed by this polyextremotolerant and morphologically plastic organism.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Terms of Use
Creative Commons Attribution-Noncommercial-ShareAlike
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DOI of Published Version
https://doi.org/10.1083/jcb.202402114
