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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...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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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. |
format | Online Article Text |
id | pubmed-5071762 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
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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