title: Nutrient supplements boost yeast transformation efficiency.
abstract: 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 107CFU per μg plasmid DNA and per 108cells 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.
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Sheng-Chun
authors.initials: SC
authors.lastname: Yu
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Alexander
authors.initials: A
authors.lastname: Dawson
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Alyssa C
authors.initials: AC
authors.lastname: Henderson
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Eloise J
authors.initials: EJ
authors.lastname: Lockyer
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Emily
authors.initials: E
authors.lastname: Read
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Gayathri
authors.initials: G
authors.lastname: Sritharan
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Marjah
authors.initials: M
authors.lastname: Ryan
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Mara
authors.initials: M
authors.lastname: Sgroi
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Pok M
authors.initials: PM
authors.lastname: Ngou
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Rosie
authors.initials: R
authors.lastname: Woodruff
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Ruifeng
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Travis
authors.initials: T
authors.lastname: Ren Teen Chia
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Yiyu
authors.initials: Y
authors.lastname: Xiang
authors.affiliation: Department of Life Sciences, South Kensington Campus, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Pietro D
authors.initials: PD
authors.lastname: Spanu
mesh_terms: D001685:Biological Factors / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry*; D061269:DNA Transformation Competence / Q000187:drug effects*; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics*
chemical_list: D001685:Biological Factors; D003470:Culture Media