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Development of a Transformation Method for Metschnikowia borealis and other CUG-Serine Yeasts

Yeasts belonging to the Metschnikowia genus are particularly interesting for the unusual formation of only two needle-shaped ascospores during their mating cycle. Presently, the meiotic process that can lead to only two spores from a diploid zygote is poorly understood. The expression of fluorescent...

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Autores principales: Gordon, Zachary B., Soltysiak, Maximillian P.M., Leichthammer, Christopher, Therrien, Jasmine A., Meaney, Rebecca S., Lauzon, Carolyn, Adams, Matthew, Lee, Dong Kyung, Janakirama, Preetam, Lachance, Marc-André, Karas, Bogumil J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6409616/
https://www.ncbi.nlm.nih.gov/pubmed/30678093
http://dx.doi.org/10.3390/genes10020078
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author Gordon, Zachary B.
Soltysiak, Maximillian P.M.
Leichthammer, Christopher
Therrien, Jasmine A.
Meaney, Rebecca S.
Lauzon, Carolyn
Adams, Matthew
Lee, Dong Kyung
Janakirama, Preetam
Lachance, Marc-André
Karas, Bogumil J.
author_facet Gordon, Zachary B.
Soltysiak, Maximillian P.M.
Leichthammer, Christopher
Therrien, Jasmine A.
Meaney, Rebecca S.
Lauzon, Carolyn
Adams, Matthew
Lee, Dong Kyung
Janakirama, Preetam
Lachance, Marc-André
Karas, Bogumil J.
author_sort Gordon, Zachary B.
collection PubMed
description Yeasts belonging to the Metschnikowia genus are particularly interesting for the unusual formation of only two needle-shaped ascospores during their mating cycle. Presently, the meiotic process that can lead to only two spores from a diploid zygote is poorly understood. The expression of fluorescent nuclear proteins should allow the meiotic process to be visualized in vivo; however, no large-spored species of Metschnikowia has ever been transformed. Accordingly, we aimed to develop a transformation method for Metschnikowia borealis, a particularly large-spored species of Metschnikowia, with the goal of enabling the genetic manipulations required to study biological processes in detail. Genetic analyses confirmed that M. borealis, and many other Metschnikowia species, are CUG-Ser yeasts. Codon-optimized selectable markers lacking CUG codons were used to successfully transform M. borealis by electroporation and lithium acetate, and transformants appeared to be the result of random integration. Mating experiments confirmed that transformed-strains were capable of generating large asci and undergoing recombination. Finally, random integration was used to transform an additional 21 yeast strains, and all attempts successfully generated transformants. The results provide a simple method to transform many yeasts from an array of different clades and can be used to study or develop many species for various applications.
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spelling pubmed-64096162019-03-26 Development of a Transformation Method for Metschnikowia borealis and other CUG-Serine Yeasts Gordon, Zachary B. Soltysiak, Maximillian P.M. Leichthammer, Christopher Therrien, Jasmine A. Meaney, Rebecca S. Lauzon, Carolyn Adams, Matthew Lee, Dong Kyung Janakirama, Preetam Lachance, Marc-André Karas, Bogumil J. Genes (Basel) Article Yeasts belonging to the Metschnikowia genus are particularly interesting for the unusual formation of only two needle-shaped ascospores during their mating cycle. Presently, the meiotic process that can lead to only two spores from a diploid zygote is poorly understood. The expression of fluorescent nuclear proteins should allow the meiotic process to be visualized in vivo; however, no large-spored species of Metschnikowia has ever been transformed. Accordingly, we aimed to develop a transformation method for Metschnikowia borealis, a particularly large-spored species of Metschnikowia, with the goal of enabling the genetic manipulations required to study biological processes in detail. Genetic analyses confirmed that M. borealis, and many other Metschnikowia species, are CUG-Ser yeasts. Codon-optimized selectable markers lacking CUG codons were used to successfully transform M. borealis by electroporation and lithium acetate, and transformants appeared to be the result of random integration. Mating experiments confirmed that transformed-strains were capable of generating large asci and undergoing recombination. Finally, random integration was used to transform an additional 21 yeast strains, and all attempts successfully generated transformants. The results provide a simple method to transform many yeasts from an array of different clades and can be used to study or develop many species for various applications. MDPI 2019-01-23 /pmc/articles/PMC6409616/ /pubmed/30678093 http://dx.doi.org/10.3390/genes10020078 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Gordon, Zachary B.
Soltysiak, Maximillian P.M.
Leichthammer, Christopher
Therrien, Jasmine A.
Meaney, Rebecca S.
Lauzon, Carolyn
Adams, Matthew
Lee, Dong Kyung
Janakirama, Preetam
Lachance, Marc-André
Karas, Bogumil J.
Development of a Transformation Method for Metschnikowia borealis and other CUG-Serine Yeasts
title Development of a Transformation Method for Metschnikowia borealis and other CUG-Serine Yeasts
title_full Development of a Transformation Method for Metschnikowia borealis and other CUG-Serine Yeasts
title_fullStr Development of a Transformation Method for Metschnikowia borealis and other CUG-Serine Yeasts
title_full_unstemmed Development of a Transformation Method for Metschnikowia borealis and other CUG-Serine Yeasts
title_short Development of a Transformation Method for Metschnikowia borealis and other CUG-Serine Yeasts
title_sort development of a transformation method for metschnikowia borealis and other cug-serine yeasts
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6409616/
https://www.ncbi.nlm.nih.gov/pubmed/30678093
http://dx.doi.org/10.3390/genes10020078
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