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Increased Agrobacterium‐mediated transformation of Saccharomyces cerevisiae after deletion of the yeast ADA2 gene

Agrobacterium tumefaciens is the causative agent of crown gall disease and is widely used as a vector to create transgenic plants. Under laboratory conditions, the yeast Saccharomyces cerevisiae and other yeasts and fungi can also be transformed, and Agrobacterium‐mediated transformation (AMT) is no...

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Autores principales: Roushan, M.R., Shao, S., Poledri, I., Hooykaas, P.J.J., van Heusden, G.P.H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9299121/
https://www.ncbi.nlm.nih.gov/pubmed/34816457
http://dx.doi.org/10.1111/lam.13605
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author Roushan, M.R.
Shao, S.
Poledri, I.
Hooykaas, P.J.J.
van Heusden, G.P.H.
author_facet Roushan, M.R.
Shao, S.
Poledri, I.
Hooykaas, P.J.J.
van Heusden, G.P.H.
author_sort Roushan, M.R.
collection PubMed
description Agrobacterium tumefaciens is the causative agent of crown gall disease and is widely used as a vector to create transgenic plants. Under laboratory conditions, the yeast Saccharomyces cerevisiae and other yeasts and fungi can also be transformed, and Agrobacterium‐mediated transformation (AMT) is now considered the method of choice for genetic transformation of many fungi. Unlike plants, in S. cerevisiae, T‐DNA is integrated preferentially by homologous recombination and integration by non‐homologous recombination is very inefficient. Here we report that upon deletion of ADA2, encoding a component of the ADA and SAGA transcriptional adaptor/histone acetyltransferase complexes, the efficiency of AMT significantly increased regardless of whether integration of T‐DNA was mediated by homologous or non‐homologous recombination. This correlates with an increase in double‐strand DNA breaks, the putative entry sites for T‐DNA, in the genome of the ada2Δ deletion mutant, as visualized by the number of Rad52‐GFP foci. Our observations may be useful to enhance the transformation of species that are difficult to transform.
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spelling pubmed-92991212022-07-21 Increased Agrobacterium‐mediated transformation of Saccharomyces cerevisiae after deletion of the yeast ADA2 gene Roushan, M.R. Shao, S. Poledri, I. Hooykaas, P.J.J. van Heusden, G.P.H. Lett Appl Microbiol Original Articles Agrobacterium tumefaciens is the causative agent of crown gall disease and is widely used as a vector to create transgenic plants. Under laboratory conditions, the yeast Saccharomyces cerevisiae and other yeasts and fungi can also be transformed, and Agrobacterium‐mediated transformation (AMT) is now considered the method of choice for genetic transformation of many fungi. Unlike plants, in S. cerevisiae, T‐DNA is integrated preferentially by homologous recombination and integration by non‐homologous recombination is very inefficient. Here we report that upon deletion of ADA2, encoding a component of the ADA and SAGA transcriptional adaptor/histone acetyltransferase complexes, the efficiency of AMT significantly increased regardless of whether integration of T‐DNA was mediated by homologous or non‐homologous recombination. This correlates with an increase in double‐strand DNA breaks, the putative entry sites for T‐DNA, in the genome of the ada2Δ deletion mutant, as visualized by the number of Rad52‐GFP foci. Our observations may be useful to enhance the transformation of species that are difficult to transform. John Wiley and Sons Inc. 2021-11-27 2022-02 /pmc/articles/PMC9299121/ /pubmed/34816457 http://dx.doi.org/10.1111/lam.13605 Text en © 2021 The Authors. Letters in Applied Microbiology published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Original Articles
Roushan, M.R.
Shao, S.
Poledri, I.
Hooykaas, P.J.J.
van Heusden, G.P.H.
Increased Agrobacterium‐mediated transformation of Saccharomyces cerevisiae after deletion of the yeast ADA2 gene
title Increased Agrobacterium‐mediated transformation of Saccharomyces cerevisiae after deletion of the yeast ADA2 gene
title_full Increased Agrobacterium‐mediated transformation of Saccharomyces cerevisiae after deletion of the yeast ADA2 gene
title_fullStr Increased Agrobacterium‐mediated transformation of Saccharomyces cerevisiae after deletion of the yeast ADA2 gene
title_full_unstemmed Increased Agrobacterium‐mediated transformation of Saccharomyces cerevisiae after deletion of the yeast ADA2 gene
title_short Increased Agrobacterium‐mediated transformation of Saccharomyces cerevisiae after deletion of the yeast ADA2 gene
title_sort increased agrobacterium‐mediated transformation of saccharomyces cerevisiae after deletion of the yeast ada2 gene
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9299121/
https://www.ncbi.nlm.nih.gov/pubmed/34816457
http://dx.doi.org/10.1111/lam.13605
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