Su Jianchen, Ouyang Haomiao, Zhao Wan, Dong Zhiyang, Jin Cheng
State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
Wei Sheng Wu Xue Bao. 2013 Jan 4;53(1):38-46.
AfMp1p is a glycosylphosphatidylinositol (GPI) anchored cell wall protein (GPI-CWP) identified in filamentous fungus Aspergillus fumigatus, which contains a specific C terminal signal for cell wall localization. Trichoderma reesei is known as a safe fungal species (GRAS) and widely used in the industry. Thus, developing of the cell-surface expression systems in T. reesei is of industrial interest.
The GPI signal from the A. fumigatus AfMp1p was fused to the C-terminal of green fluorescent protein (GFP) and transformed into T. reesei. The optimization of T. reesei transformation and the expression profiles of the GFP were investigated in detail. The cellular location of the GFP fusion protein was detected.
Fluorescent image analysis and Western blot analysis indicate that the GFP fusion protein locates on the cell wall of T. reesei.
According to these results, the GPI signal from A. fumigatus can be recognized in T. reesei and the expression system constructed in this study can be used to express heterogeneous protein in the cell wall of T. reesei.
AfMp1p是在丝状真菌烟曲霉中鉴定出的一种糖基磷脂酰肌醇（GPI）锚定细胞壁蛋白（GPI-CWP），其含有用于细胞壁定位的特定C末端信号。里氏木霉是一种安全的真菌物种（公认安全），并在工业中广泛使用。因此，开发里氏木霉的细胞表面表达系统具有工业价值。
将来自烟曲霉AfMp1p的GPI信号与绿色荧光蛋白（GFP）的C末端融合，并转化到里氏木霉中。详细研究了里氏木霉转化的优化以及GFP的表达谱。检测了GFP融合蛋白的细胞定位。
荧光图像分析和蛋白质印迹分析表明，GFP融合蛋白定位于里氏木霉的细胞壁上。
根据这些结果，烟曲霉的GPI信号在里氏木霉中可被识别，并且本研究构建的表达系统可用于在里氏木霉的细胞壁中表达异源蛋白。
