High Efficiency Transformation by Electroporation of Yarrowia lipolytica

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 pret...

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Veröffentlicht in:The journal of microbiology 2011, 49(3), , pp.469-472
Hauptverfasser: 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
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Sprache:eng
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Zusammenfassung: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.
ISSN:1225-8873
1976-3794
DOI:10.1007/s12275-011-0433-6