Title High Efficiency Transformation by Electroporation of Yarrowia lipolytica
Author Jia-Hung Wang, Wenpin Hung, and Shu-Hsien Tsai*
Address Material and Chemical Research Laboratories, Industrial Technology Research Institute, Hsinchu 300, Taiwan
Bibliography Journal of Microbiology, 49(3),469-472, 2011,
Key Words dithiothreitol, electroporation, lithium acetate, transformation, Yarrowia lipolytica
Abstract 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 1010/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/μg for integrative linear DNA. With our modifications, the electroporation procedures became a very efficient and reliable tool for Y. lipolytica transformation.