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Efficient transformation of the entomopathogenic fungus Lecanicillium muscarium by electroporation of germinated conidia |
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| Authors: | Sergey Timofeev Alexander Tsarev Igor Senderskiy Eugene Rogozhin Galina Mitina Sergey Kozlov Viacheslav Dolgikh |
| Affiliation: | 1. Laboratory of Microbiological Control, All-Russian Institute for Plant Protection, 196608, Podbelskogo 3, St. Petersburg, Pushkin, Russia;2. Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Miklukho-Maklaya 16/10, Moscow, Russia;3. Gause Institute of New Antibiotics, 119021, Bolshaya Pirogovskaya 11, Moscow, Russia |
| Abstract: | All experiments on genetic modifications of Lecanicillium species performed so far used laborious PEG-mediated protoplast transformation and Agrobacterium tumefaciens-mediated transformation. In this study we demonstrated that simple and effective method of electroporation of germinated conidia can be used for transformation of this fungus. Electroporation of L. muscarium (strain VL 72) germinated conidia by the pBARGPE1 vector harboring an eGFP gene, showed a transformation efficiency 65.7 ± 1.4 phosphinothricin-resistant colonies per 2.5 μg of linearized plasmid DNA and expression of fluorescent protein without affecting fungal growth and virulence. By analysis of GFP-expressing isolates by fluorescent microscopy and immunoblotting, we found that the Aspergillus nidulans PgpdA promoter and trpC terminator in the pBARGPE1 vector provide an effective synthesis of heterologous proteins in L. muscarium cells. |
| Keywords: | Corresponding author GFP Heterologous expression |
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