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Nutrient supplements boost yeast transformation efficiency

Efficiency of yeast transformation is determined by the rate of yeast endocytosis. The aim of this study was to investigate the effect of introducing amino acids and other nutrients (inositol, adenine, or p-aminobenzoic acid) in the transformation medium to develop a highly efficient yeast transform...

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Autores principales: Yu, Sheng-Chun, Dawson, Alexander, Henderson, Alyssa C., Lockyer, Eloise J., Read, Emily, Sritharan, Gayathri, Ryan, Marjah, Sgroi, Mara, Ngou, Pok M., Woodruff, Rosie, Zhang, Ruifeng, Ren Teen Chia, Travis, Liu, Yu, Xiang, Yiyu, Spanu, Pietro D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5071762/
https://www.ncbi.nlm.nih.gov/pubmed/27760994
http://dx.doi.org/10.1038/srep35738
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author Yu, Sheng-Chun
Dawson, Alexander
Henderson, Alyssa C.
Lockyer, Eloise J.
Read, Emily
Sritharan, Gayathri
Ryan, Marjah
Sgroi, Mara
Ngou, Pok M.
Woodruff, Rosie
Zhang, Ruifeng
Ren Teen Chia, Travis
Liu, Yu
Xiang, Yiyu
Spanu, Pietro D.
author_facet Yu, Sheng-Chun
Dawson, Alexander
Henderson, Alyssa C.
Lockyer, Eloise J.
Read, Emily
Sritharan, Gayathri
Ryan, Marjah
Sgroi, Mara
Ngou, Pok M.
Woodruff, Rosie
Zhang, Ruifeng
Ren Teen Chia, Travis
Liu, Yu
Xiang, Yiyu
Spanu, Pietro D.
author_sort Yu, Sheng-Chun
collection PubMed
description Efficiency of yeast transformation is determined by the rate of yeast endocytosis. The aim of this study was to investigate the effect of introducing amino acids and other nutrients (inositol, adenine, or p-aminobenzoic acid) in the transformation medium to develop a highly efficient yeast transformation protocol. The target of rapamycin complex 1 (TORC1) kinase signalling complex influences the rate of yeast endocytosis. TORC signaling is induced by amino acids in the media. Here, we found that increasing the concentration of amino acids and other nutrients in the growth media lead to an increase yeast transformation efficiency up to 10(7) CFU per μg plasmid DNA and per 10(8) cells with a 13.8 kb plasmid DNA. This is over 130 times that of current published methods. This improvement may facilitate more efficient experimentation in which transformation efficiency is critical, such as yeast two-hybrid screening.
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spelling pubmed-50717622016-10-26 Nutrient supplements boost yeast transformation efficiency Yu, Sheng-Chun Dawson, Alexander Henderson, Alyssa C. Lockyer, Eloise J. Read, Emily Sritharan, Gayathri Ryan, Marjah Sgroi, Mara Ngou, Pok M. Woodruff, Rosie Zhang, Ruifeng Ren Teen Chia, Travis Liu, Yu Xiang, Yiyu Spanu, Pietro D. Sci Rep Article Efficiency of yeast transformation is determined by the rate of yeast endocytosis. The aim of this study was to investigate the effect of introducing amino acids and other nutrients (inositol, adenine, or p-aminobenzoic acid) in the transformation medium to develop a highly efficient yeast transformation protocol. The target of rapamycin complex 1 (TORC1) kinase signalling complex influences the rate of yeast endocytosis. TORC signaling is induced by amino acids in the media. Here, we found that increasing the concentration of amino acids and other nutrients in the growth media lead to an increase yeast transformation efficiency up to 10(7) CFU per μg plasmid DNA and per 10(8) cells with a 13.8 kb plasmid DNA. This is over 130 times that of current published methods. This improvement may facilitate more efficient experimentation in which transformation efficiency is critical, such as yeast two-hybrid screening. Nature Publishing Group 2016-10-20 /pmc/articles/PMC5071762/ /pubmed/27760994 http://dx.doi.org/10.1038/srep35738 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Yu, Sheng-Chun
Dawson, Alexander
Henderson, Alyssa C.
Lockyer, Eloise J.
Read, Emily
Sritharan, Gayathri
Ryan, Marjah
Sgroi, Mara
Ngou, Pok M.
Woodruff, Rosie
Zhang, Ruifeng
Ren Teen Chia, Travis
Liu, Yu
Xiang, Yiyu
Spanu, Pietro D.
Nutrient supplements boost yeast transformation efficiency
title Nutrient supplements boost yeast transformation efficiency
title_full Nutrient supplements boost yeast transformation efficiency
title_fullStr Nutrient supplements boost yeast transformation efficiency
title_full_unstemmed Nutrient supplements boost yeast transformation efficiency
title_short Nutrient supplements boost yeast transformation efficiency
title_sort nutrient supplements boost yeast transformation efficiency
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5071762/
https://www.ncbi.nlm.nih.gov/pubmed/27760994
http://dx.doi.org/10.1038/srep35738
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