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Novel blastospore-based transformation system for integration of phosphinothricin resistance and green fluorescence protein genes into Beauveria bassiana

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Abstract

A novel system was developed for efficient transformation of the fungal biocontrol agent Beauveriabassiana. Competent blastospores were prepared and stored in LiAc- and glycerol-inclusive suspension at −76 °C for sequential use in transformation. The system was successfully applied to integrating phosphinothricin resistance gene bar and enhanced green fluorescence protein gene egfp into B. bassiana via blastospore absorption of a plasmid vectoring bar and egfp. A frequency of 24 transformants per microgram of DNA was achieved. The blastospore-based transformation system has proven to be very convenient and would be highly potential for use in genetic manipulation of B. bassiana and other filamentous species.

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Acknowledgements

Punt PJ (TNO Nutrition and Food Research Institute, Zeist, The Netherlands) and Sun ZX (China National Rice Research Institute, Hangzhou, China) are thanked for providing plasmids pAN52-1N and pDS-1, respectively. Funding of this study was provided by the National ‘Program 973’ of the Ministry of Science and Technology of China (2003CB114203), the Natural Science Foundation of China (30571250), and Zhejiang Provincial R&D Program (2003G10038).

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Authors and Affiliations

  1. Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, 310029, People’s Republic of China

    Sheng-Hua Ying & Ming-Guang Feng

  2. Institute of Insect Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, 310029, People’s Republic of China

    Ming-Guang Feng

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  1. Sheng-Hua Ying
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  2. Ming-Guang Feng
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Correspondence to Ming-Guang Feng.

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Ying, SH., Feng, MG. Novel blastospore-based transformation system for integration of phosphinothricin resistance and green fluorescence protein genes into Beauveria bassiana . Appl Microbiol Biotechnol 72, 206–210 (2006). https://doi.org/10.1007/s00253-006-0447-x

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  • DOI: https://doi.org/10.1007/s00253-006-0447-x

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