Ying Sheng-Hua, Feng Ming-Guang
Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, 310029, People's Republic of China.
Institute of Insect Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, 310029, People's Republic of China.
Appl Microbiol Biotechnol. 2006 Aug;72(1):206-210. doi: 10.1007/s00253-006-0447-x. Epub 2006 Apr 26.
A novel system was developed for efficient transformation of the fungal biocontrol agent Beauveria bassiana. Competent blastospores were prepared and stored in LiAc- and glycerol-inclusive suspension at -76 degrees 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.
开发了一种用于高效转化真菌生防菌球孢白僵菌的新系统。制备了感受态芽生孢子，并将其保存在含有LiAc和甘油的悬浮液中，于-76℃下保存，以便在转化中依次使用。通过芽生孢子吸收携带bar和egfp的质粒载体，该系统成功地将膦丝菌素抗性基因bar和增强型绿色荧光蛋白基因egfp整合到球孢白僵菌中。实现了每微克DNA有24个转化体的频率。基于芽生孢子的转化系统已被证明非常方便，在球孢白僵菌和其他丝状物种的遗传操作中具有很高的应用潜力。
