An improved method for Beauveria bassiana transformation using phosphinothricin acetyltransferase and green fluorescent protein fusion gene as a selectable and visible marker.
An improved transformation method for the biocontrol agent, Beauveria bassiana, was developed. For convenience of transformation selection and detection, the coding regions of the genes for phosphinothricin acetyltransferase and green fluorescent protein were fused and an expression vector, pBFT, carrying this fusion was constructed. Under optimum conditions, over 60 transformants microg(-1) plasmid DNA were obtained. B. bassiana conidia frozen 1 month at -80 degrees C were fully competent for transformation. The method was significantly less laborious and more rapid than current methods for B. bassiana. The bar::egfp provides a selectable and visible marker which may expedite future genetic engineering of this fungus.
Jin
Kai
K
Key Laboratory of Biotechnology and Crop Quality Improvement of Ministry of Agriculture of China, Biotechnology Research Center, Southwest University, Chongqing 400716, People's Republic of China.
Zhang
Yongjun
Y
Luo
Zhibing
Z
Xiao
Yuehua
Y
Fan
Yanhua
Y
Wu
Di
D
Pei
Yan
Y
D000123:Acetyltransferases / Q000378:metabolism*; D052982:Beauveria / Q000201:enzymology* / Q000235:genetics*; D001709:Biotechnology / Q000379:methods*; D004560:Electricity; D005615:Freezing; D005819:Genetic Markers; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D015203:Reproducibility of Results; D013172:Spores, Fungal / Q000166:cytology / Q000502:physiology; D014170:Transformation, Genetic
D005819:Genetic Markers; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D000123:Acetyltransferases; C056243:phosphinothricin N-acetyltransferase