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多芽游动酵母 Aureobasidium pullulans 的化学转化。

Chemical transformation of the multibudding yeast, Aureobasidium pullulans.

机构信息

Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA.

Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, NC, USA.

出版信息

J Cell Biol. 2024 Oct 7;223(10). doi: 10.1083/jcb.202402114. Epub 2024 Jun 27.

DOI:10.1083/jcb.202402114
PMID:38935076
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11211067/
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.

摘要

出芽短梗霉是一种普遍存在的多态性黑酵母，具有工业和农业应用价值。它最近在细胞生物学家中引起了关注，因为它的增殖方式不同寻常，多核酵母细胞在一个细胞周期内产生多个芽。在这里，我们将化学转化方法与基因组靶向同源重组相结合，仅在 3 天内就获得了约 60 个转化体/μg 的 DNA。该方案简单、廉价，且不需要特殊设备。我们还描述了带有密码子优化的绿色和红色荧光蛋白的载体，用于出芽短梗霉，并使用这些工具探索新的细胞生物学。表达细胞质和核标记的菌株的定量成像表明，尽管类似体积的细胞中核数差异很大，但总核体积与细胞体积呈显著的 70 倍大小范围的比例关系。这里描述的方案和工具扩展了出芽短梗霉生物学家的工具包，并将帮助研究人员解决这个多极端耐受和形态可塑性生物体带来的许多其他难题。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/9a98009ed254/JCB_202402114_FigS4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/c5b10b794624/JCB_202402114_Fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/8246b2fa8a82/JCB_202402114_FigS1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/a8fa233ded55/JCB_202402114_Fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/91f413c3da48/JCB_202402114_FigS2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/37b20a927d1e/JCB_202402114_Fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/7c62d53559ed/JCB_202402114_FigS3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/42b78b386f98/JCB_202402114_Fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/14339620b1f1/JCB_202402114_Fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/566bd57e9be7/JCB_202402114_Fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/9a98009ed254/JCB_202402114_FigS4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/c5b10b794624/JCB_202402114_Fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/8246b2fa8a82/JCB_202402114_FigS1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/a8fa233ded55/JCB_202402114_Fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/91f413c3da48/JCB_202402114_FigS2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/37b20a927d1e/JCB_202402114_Fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/7c62d53559ed/JCB_202402114_FigS3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/42b78b386f98/JCB_202402114_Fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/14339620b1f1/JCB_202402114_Fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/566bd57e9be7/JCB_202402114_Fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bcb/11211067/9a98009ed254/JCB_202402114_FigS4.jpg

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