丝状真菌蛋白质组分析方法的建立  

Establishment of Analytical Method in Filamentous fungal Proteome

作　　者：李锐[1] 贺亮[2] 吴俐勤[1] 

机构地区：[1]浙江省农业科学院农产品质量标准研究所,浙江杭州310021 [2]上海医药工业研究院,国家(上海)新药安全评价研究中心,上海201203

出　　处：《华北农学报》2012年第B12期93-96,共4页Acta Agriculturae Boreali-Sinica

基　　金：国家“863”项目(2006AA020504)

摘　　要：旨在建立丝状真菌蛋白质组的分析方法,对丝状真菌蛋质白学进行初步探索。使用液氮冷冻研磨、蜗牛酶消化、尿素裂解、硫酸铵沉淀预分级和二维液相色谱分离相结合的蛋白质组研究方法进行丝状真菌的蛋白质组分析;建立的丝状真菌蛋白提取分级分离方法有效的提高了蛋白的分离效率和提取数量,可以应用在丝状真菌蛋白质组研究中,共鉴定了396个蛋白,包括了参与各种细胞生理过程的蛋白,为进一步开展丝状真菌蛋白质组的研究打下了良好的基础。研究中建立的蜗牛酶消化破壁、尿素裂解的蛋白提取方法,可以显著提高丝状真菌胞浆蛋白的鉴定数量,特别是对于那些功能重要但丰度低的蛋白,适合于丝状真菌蛋白质组的研究。To develop the methods of Filamentous fungal proteome analysis for preliminary research.Liquid nitrogen grinding,snailase digestion,urea lysis and ammonium sulfate fractionation,two-dimensional liquid chromatography separation methods was established for application in Filamentous fungal proteome analysis.The method increases the efficiency of extraction and separation of the protein for applying to Filamentous fungal proteome analysis.396 T.rubrum proteins were unambiguously identified involving in major biological processes and this experiment was a basis for further fungal proteomic research.These results suggested that this protein extraction,prefraction and separation methods were effective and sensitive for detection interest proteins in Filamentous fungal proteome analysis,especially important functional low abundance proteins.

关 键 词：丝状真菌 蜗牛酶 尿素裂解 蛋白质组 

分 类 号：S855.4[农业科学—临床兽医学]