Novel blastospore-based transformation system for integration of phosphinothricin resistance and green fluorescence protein genes into Beauveria bassiana.
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.
Ying
Sheng-Hua
SH
Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, 310029, People's Republic of China.
Feng
Ming-Guang
MG
Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, 310029, People's Republic of China. mgfeng@zju.edu.cn.|Institute of Insect Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, 310029, People's Republic of China. mgfeng@zju.edu.cn.
D000613:Aminobutyrates / Q000494:pharmacology*; D000935:Antifungal Agents / Q000494:pharmacology; D032661:Cordyceps / Q000187:drug effects / Q000235:genetics*; D026141:Drug Resistance, Multiple, Fungal / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006999:Hypocreales; D010957:Plasmids / Q000235:genetics; D013172:Spores, Fungal / Q000235:genetics; D014170:Transformation, Genetic
D000613:Aminobutyrates; D000935:Antifungal Agents; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C003121:phosphinothricin