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KMID : 0880220110490030469
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Journal of Microbiology
2011 Volume.49 No. 3 p.469 ~ p.472
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High efficiency transformation by electroporation of Yarrowia lipolytica
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Jia-Hung Wang
Wenpin Hung
Shu-Hsien Tsai
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Abstract
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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 F. lipolytica, we report here that pretreatment of F. 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 F. lipolytica that will be able to yield an efficiency of 2.1 × 104 transformants/ug for integrative linear DNA. With our modifications, the electroporation procedures became a very efficient and reliable tool for F. lipolytica transformation.
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KEYWORD
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dithiothreitol, electroporation, lithium acetate, transformation, Yarrowia lipolytica
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