Yang Yun-Jung, Lee Inhyung
Department of Foods and Nutrition, Kookmin University, Seoul 132-706, Korea.
J Microbiol Biotechnol. 2008 Apr;18(4):754-8.
Agrobacterium tumefaciens-mediated transformation (ATMT) was successfully applied to Monascus ruber. The optimum cocultivation time was 84 h with an efficiency of 900 to 1,000 transformants when 1x106 spores were used with the same volume of bacteria. The stability of transformants was over 98% after five generations. When M. ruber was transformed with A. tumefaciens YL-63 containing the green fluorescent protein gene (egfp), the green fluorescent signal was observed throughout hyphae, confirming expression of the gene. This efficient transformation and expression system of M. ruber by ATMT will facilitate the study of this fungus at a molecular genetic level.
根癌农杆菌介导的转化(ATMT)已成功应用于红曲霉。当使用1×10⁶个孢子与相同体积的细菌时,最佳共培养时间为84小时,转化效率为900至1000个转化体。五代后转化体的稳定性超过98%。当用含有绿色荧光蛋白基因(egfp)的根癌农杆菌YL-63转化红曲霉时,在整个菌丝中观察到绿色荧光信号,证实了该基因的表达。这种通过ATMT对红曲霉进行高效转化和表达的系统将有助于在分子遗传水平上对这种真菌进行研究。