Nutrient supplements boost yeast transformation efficiency

Sci Rep. 2016 Oct 20:6:35738. doi: 10.1038/srep35738.

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 107 CFU per μg plasmid DNA and per 108 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Factors / metabolism
  • Culture Media / chemistry*
  • DNA Transformation Competence / drug effects*
  • Saccharomyces cerevisiae / drug effects*
  • Saccharomyces cerevisiae / genetics*

Substances

  • Biological Factors
  • Culture Media