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

作　　者：张宇[1] 贺丹[1] 王爽[2] 张艳华[1,3] 李涵[1] 王丽[1] 

机构地区：[1]吉林大学白求恩医学院病原生物学系,长春130021 [2]吉林大学第二医院皮肤科,长春130041 [3]吉林大学植物科学学院生物学系,长春130062

出　　处：《中国生物制品学杂志》2010年第3期313-316,共4页Chinese Journal of Biologicals

基　　金：国家自然科学基金项目资助(30770120)

摘　　要：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.

关 键 词：烟曲霉 分泌蛋白质 双向凝胶电泳 TCA 丙酮 

分 类 号：R379[医药卫生—病原生物学] R392.33[医药卫生—基础医学]