Wang Jieping, Wang Jiaxu, Liu Fan, Pan Cangsang
School of Life Science, Xiamen University, Xiamen 361005, China.
Sheng Wu Gong Cheng Xue Bao. 2010 May;26(5):630-4.
The main aim of this study was to transform the enhanced green fluorescent protein gene (egfp) into biocontrol fungus Paecilomyces lilacinus strain 9410. We constructed the expression vector pUPNGT of the fusion gene nptII-egfp using pcDNA3.1(-) as a helper plasmid. The egfp gene was then transformed into P. lilacinus strain 9410 via Agrobacterium tumefaciens-mediated transformation. PCR and Southern blotting analysis showed that the egfp gene was integrated into the genomes of the tested transformants and the integration manner was single-copy. The transformants could generate green fluorescence when they were excited by 488 nm blue laser. These results indicated that the egfp gene had been successfully transformed into P. lilacinus 9410 and expressed in the tested transformants. Our work may provide a new approach to assess environmental safety and practical biocontrol efficacy ofP. lilacinus under different conditions.
本研究的主要目的是将增强型绿色荧光蛋白基因（egfp）导入生防真菌淡紫拟青霉9410菌株。我们以pcDNA3.1(-)作为辅助质粒构建了融合基因nptII-egfp的表达载体pUPNGT。然后通过根癌农杆菌介导的转化将egfp基因导入淡紫拟青霉9410菌株。PCR和Southern杂交分析表明，egfp基因已整合到测试转化子的基因组中，且整合方式为单拷贝。当用488 nm蓝色激光激发时，转化子能够产生绿色荧光。这些结果表明，egfp基因已成功导入淡紫拟青霉9410并在测试转化子中表达。我们的工作可能为评估淡紫拟青霉在不同条件下的环境安全性和实际生防效果提供一种新方法。
