2010•Acta Scientiarum Naturalium Universitatis SunyatseniRequires access
A Highly Efficient Protocol for Transformation of Saccharomyces cerevisiae and Pichia pastoris by Electroporation
Zhiwu Xu, Ying Zhang, Zhixue Wang, Yan Cao, Cui Liang, Sheng Li, Qiuyun Liu
Open publisher page 5 citations
Abstract
Electroporation is widely used for highly efficient transformation of Saccharomyces cerevisiae.As many strains degenerate over time or over environmental exposure,their electroporation efficiencies could drop markedly by 2 to 3 orders of magnitude.Here we describe a modified electroporation procedure which included single stranded carrier DNA for electroporation as well as a post-electroporation growth period in YPD medium,which gave rise to 13 fold enhancement in transformations than an optimized lithium acetate(LiAc) and dithiothreitol(DTT) Pretreatment protocol previously reported.An enhancement of 114 fold on Pichia pastoris transformations has been observed using the modified procedure.The modified method enhanced the transformation efficiencies of some degenerated S.cerevisiae strains by nearly 100 fold.It will enable molecular manipulations in all kinds of S.cerevisiae and P.pastoris strains by allowing propagations of all sorts of DNA libraries made from minute samples.