High efficiency transformation by electroporation of <Emphasis Type="Italic">Yarrowia lipolytica</Emphasis> |
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Authors: | Jia-Hung Wang Wenpin Hung Shu-Hsien Tsai |
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Institution: | (1) Department of Biotechnology and Food Microbiology, Wroclaw University of Environmental and Life Sciences, Norwida 25, 50-375 Wroclaw, Poland |
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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 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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