High Efficiency Transformation by Electroporation of Yarrowia lipolytica
2011
Wang, Jia-Hung, Industrial Technology Research Institute, Hsinchu, Taiwan | Hung, Wenpin, Industrial Technology Research Institute, Hsinchu, Taiwan | Tsai, Shu-Hsien, Industrial Technology Research Institute, Hsinchu, Taiwan
Yarrowia lipolytica was usually transformed by heat shock, but linearized integrative vectors always resulted in a low transformation efficiency when electroporation was used. To develop a high efficiency integrative transformation method by electroporation of Y. lipolytica, we report here that pretreatment of Y. lipolytica with 150 mM LiAc for 1 h before electroporation will approximately 30-fold of increase transformation efficiency. A cell concentration of 10∨10/ml and instrument settings of 1.5 kV will generate the highest transformation efficiencies. We have developed a procedure to transform Y. lipolytica that will be able to yield an efficiency of 2.1×10⁴ transformants/mu g for integrative linear DNA. With our modifications, the electroporation procedures became a very efficient and reliable tool for Y. lipolytica transformation.
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