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目的比较3种丝状真菌烟曲霉分泌蛋白质的提取方法,并通过双向凝胶电泳对提取的蛋白质进行分离。方法利用TCA沉淀、丙酮沉淀及TCA-丙酮沉淀法分别提取烟曲霉的分泌蛋白质,SDS-PAGE比较其结果,初步确定最佳提取方法,并利用双向凝胶电泳分离提取的蛋白质。结果TCA-丙酮沉淀法提取的蛋白质浓度最高,种类和数量最多,双向凝胶电泳显示蛋白质点聚合较好,无非特异的条带和杂质,背景透明。结论TCA-丙酮沉淀法提取的烟曲霉分泌蛋白质图谱较好,为丝状真菌分泌蛋白质的研究奠定了基础。
Objective To compare the extraction methods of three kinds of filamentous fungi, Aspergillus fumigatus, and to separate the extracted proteins by two-dimensional gel electrophoresis. Methods The secretory proteins of Aspergillus fumigatus were extracted by TCA precipitation, acetone precipitation and TCA-acetone precipitation respectively. The results were compared by SDS-PAGE and the best extraction method was determined. The extracted proteins were separated by two-dimensional gel electrophoresis. Results TCA-acetone precipitation had the highest protein concentration and the largest number of species. The two-dimensional gel electrophoresis showed that the protein spots polymerized well, without specific bands and impurities, and the background was clear. Conclusion The protein profile of Aspergillus fumigatus extracted by TCA-acetone precipitation is good, which lays the foundation for the study of the secretion of protein by filamentous fungi.