Jin Kai, Zhang Yongjun, Luo Zhibing, Xiao Yuehua, Fan Yanhua, Wu Di, Pei Yan
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.
Biotechnol Lett. 2008 Aug;30(8):1379-83. doi: 10.1007/s10529-008-9713-6. Epub 2008 Apr 15.
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.
开发了一种用于生防菌球孢白僵菌的改进转化方法。为便于转化筛选和检测，将草丁膦乙酰转移酶基因和绿色荧光蛋白基因的编码区融合，并构建了携带该融合基因的表达载体pBFT。在最佳条件下，每微克质粒DNA可获得60多个转化子。在-80℃冷冻1个月的球孢白僵菌分生孢子完全具备转化能力。该方法比目前用于球孢白僵菌的方法显著省力且更快速。bar::egfp提供了一个可选择且可见的标记，这可能会加快该真菌未来的基因工程进程。
