烟曲霉分泌蛋白质提取方法的比较

Comparison of Methods for Extraction of Protein Secreted from Aspergillus fumigatus

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 three methods for extraction of protein secreted from Aspergillus fumigatus and separate the extracted protein by two dimensional gel electrophoresis（2-DE）.Methods The protein secreted from Aspergillus fumigatus was extracted by TCA precipitation,acetone precipitation and TCA-acetone precipitation respectively,and the results were compared by SDS-PAGE to optimize the method for extraction.The extracted protein was separated by 2-DE.Results Compared with those by TCA precipitation and acetone precipitation,the protein extracted by TCA-acetone precipitation was at high concentration and in more kinds and large quantity.2-DE showed good spot aggregation of protein and transparent background,while no non-specific band or foreign matters were observed.Conclusion Clear proteomic spectrum was obtained by extraction of Aspergillus fumigatus-secreted protein by TCA-acetone precipitation,which laid a foundation of study on Aspergillus fumigatus-secreted protein.